Home Home | Template

LOADING

safety

Overview

This document describes the safety of our project and certain steps followed in and outside the laboratory to ensure this all along its development.

Safety in research refers to the practices and principles that one shall follow to prevent exposure to biological organisms, products, or material; their accidental release; or any potential harm they may cause to any individual, external organism or the environment.

Safety Skills

Hazard: a potential source of danger to a vulnerable target.

It involves:

Comprehension of the descriptive terms regarding hazards.

Knowing how to interpret chemical labels, Safety Data Sheets, and other safety information resources.
To evaluate risks from exposure to hazards: Consider both products and circumstances.
How can the risk be prevented or controlled?
Design experiments to minimize any hazards exposure and risks.

Handle, store, and dispose wastes properly.

Review and learn from past incidents.
Fires, injuries, spills.

How to react correctly to an emergency.

Clear labeling and accessible location of safety devices (showers, eye washes, fire extinguishers, spill kits)

Understandable and straightforward emergency plan including: emergency phone numbers, alarms,and escape routes.

When designing and developing our project we tried to minimize the risks as much as possible and successfully comply with all of iGEM's safety and security policies. The next paragraphs describe our laboratory’s safety measures, and how and why we implemented different biosafety and biosecurity requirements during the development of our project.

During the whole experimental stage our team worked in the laboratories of Tecnologico de Monterrey campus Chihuahua found in the School of Engineering building, which fall under Biosafety Level 2. At all times our team was supervised by any of our instructors (Ph.D. González, Ph.D. Oseguera) or by the laboratory managers (M.S. Apodaca, Ms. Hernández).

Before starting our work in the lab, all team members had to pass different safety tests and receive training. Those tests included an introduction to laboratory equipment (i.e. centrifuge, plate reader, gel chamber, autoclaves, biosafety cabinets) and covered the use of chemicals (i.e. chloroform, phenol, staining reagents). We made sure all of our team members (Biotechnological Engineering students) received the proper training and certifications , regardless of their area (Human Practices, Entrepreneurship) foreseeing any situation out of the ordinary which could compromise their safety. That way all of the team was prepared for any incident.

Safety Courses

Prior to starting the lab work. all team members completed several courses provided by the Public Health Agency of Canada web page, and were certified by them. ²

Biosecurity Plan
General Safety for Containment Labs
Containment Level 2 Operational Practices
Introduction to Biosafety
Introduction to Biosecurity
Personal Protective Equipment
Autoclaves
Microbiology Overview
Pathogen Risk Assessment
Biological Safety Cabinets
Decontamination in the Laboratory
Chemical Disinfectants

◀ ▶

Safety Guidelines

Working in a laboratory comes with potential risks, so we followed certain basic laboratory rules to ensure individual and collective safety.

Personal Protective Equipment

We made sure to always comply with the personal protective equipment (PPE) required for safe practices inside the laboratory. This includes proper clothing: long pants that cover the ankles, close-toed shoes, and long hair tied back. Apart from this a lab coat, nitrile gloves, and masks were compulsory.

The personal protective equipment was always removed when leaving the lab, as it should not be taken outside unless it's appropriately packed and sealed.. The disposable equipment was properly discarded, and the rest was arranged in its designed place.

Additional Hygiene Measures

We thoroughly washed our hands when entering the lab and after leaving it. Additionally we always disinfected our hands with 70% ethanol, before and after putting on the gloves.

Safe Spaces

Another safety policy we followed in the lab was to keep our personal belongings away from the lab when handling infectious materials to prevent any contamination in both ways.

Equipment

Autoclaves

Our lab is equipped with 3 autoclaves: Tuttnauer 2340M, All American pressure steam sterilizer 25x, 110v and Autoclave vertical AESA, CV3. The autoclave is a strong heated container using high pressures and temperatures for steam sterilization.

Biosafety Cabinets

Our lab is equipped with 4 biosafety cabinets, which prevent the samples we work with from being contaminated by either ourselves or external agents, and from contaminating our surroundings and any individual in the laboratory. As part of our protocols, we disinfected the cabinet with 70% ethanol and turned the UV light for 15 minutes minimum before using the biosafety cabinets. When introducing our hands or any other material we sprayed them thoroughly with 70% ethanol. Finally, when done using the biosafety cabinets, we made sure to clean them and let the UV light sterilize them for another 15 minutes.

Burners: For the creation of sterile zones (sterile halo)

We count with two types of burners in our lab: Meker-Fisher burners and Bunsen burners, which are used to keep the area as sterile as possible when working in an open bench. After disinfecting the working area with 70% ethanol, we used a burner to create a sterile zone to prevent contamination of the samples we were working with.

Bacteria Incinerator

Our laboratory counts with a bacteria incinerator, which was used to sterilize the inoculation loops in order to inoculate inside the biosafety cabinet.

Laboratory Management

Access

The laboratories we worked in are categorized as Containment Level 2 Laboratories. These laboratories have lockable doors and access is restricted to Biotechnology alumni and Faculty teachers. Students must complete a safety course before being granted access to any of the laboratories, as all of us did.

Waste Disposal

Laboratory waste is usually classified into four types depending on their nature, and each of them should be disposed accordingly in order to avoid accidents or contamination. Non-solid biological wastes, such as fluids, go in a yellow hermetic bottle, while solid biological wastes should be disposed in a yellow bag. Sharp objects must be disposed in a thick red plastic bottle. Objects and materials that were in contact with bacteria or any kind of infectious agent should be disposed in a red bag. These last two options were how the waste generated during our experiments was disposed of, since we only worked with waste of that nature.

Other Risk Assessments

Choosing a non-pathogenic chassis

Tec-Chihuahua values safe laboratory practices and recognizes the importance of minimizing risks to the team, our community and the environment. This is why we decided to use a non-pathogenic chassis. We chose three strains of E. coli : DH5α for vector cloning and BL21 DE3 and Shuffle T7 for protein expression. These strains are classified as Risk Group 1 organisms and generally do not cause disease in healthy adult humans nor represent a risk to the environment.

Despite the low level of risk associated with them, all protocols involving these organisms were performed in a biosafety cabinet while wearing PPE.

Working with safe parts

All parts required for the development of our project are either on iGEM’s Registry or are protein coding genes from plants which aren’t toxic to humans, animals or the environment in any way.

Verticillium dahliae

Our project involved direct work with Verticillium wilt, a fungal plant disease caused by the facultative parasite ascomycete Verticillium dahliae. Getting to know the main characteristics of this fungus and developing the right containment methods to avoid accidental release, was indispensable for the correct development of our project.

According to DSMZ, Verticillium dahliae falls within Risk Group 1 representing a low potential risk. ¹ Biological safety and biocontainment practices followed to ensure proper management of the fungus include the following:

Wearing a laboratory coat, face masks, gloves and eye protection when working with the mould cultures
Regular cleaning of laboratory attire and personal clothing
Washing our hands after working with the pathogens and before leaving the laboratory
Decontaminating work surfaces after completion of work and after any spill of the fungus
Leaving our personal belongings away from the lab
Regular decontamination of laboratory equipment and materials.

It should be noted that the handling of this fungus was done by lighting a burner in the working area to create a sterile zone. An individual incubator was designated solely for the storage of our fungi. In this way we minimized any contamination risk.

The next link, DSMZ presents more information about the characteristics of V. dahliae: https://www.dsmz.de/catalogues/details/culture/DSM-63084.html?tx_dsmzresources_pi5%5BreturnPid%5D=304

References

DSMZ - German Collection of Microorganisms and Cell Cultures . (2019). Verticillium dahliae. June 2019, de Leibniz Institute Sitio web: https://www.dsmz.de/collection/catalogue/details/culture/DSM-63084?tx_dsmzresources_pi5%5BreturnPid%5D=304
Public Health Department. (2018). Laboratory Biosafety and Biosecurity Courses. June 2019, de Government of Canada Sitio web: https://training-formation.phac-aspc.gc.ca/?lang=en

Team:Tec-Chihuahua/Safety