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From 2011.igem.org

The objective of this project is to develop a rubber degrading bacteria, and as part of it a biosensor for this product detection, using genes from bacterias that degrade rubber as part of their natural metabolism and assamble them into an E. coli chassis. We want to obtain a characterized biosensor and a synthetic enginereed bacteria that will help us reduce rubber and its derivates such as latex gloves, preservatives and tires.

Biological sciences are a great tool for innovation and improvement of human society; nevertheless when the biosafety parameters are ignored, this area of research can become quite dangerous. We always have that idea present while working at the laboratory. We follow the rules stated at the Good Laboratory Practice (GLP) and according to the safety guidelines established by the Instituto de Fisiología Celular (IFC), part of the Universidad Nacional Autónoma de México (UNAM). Furthermore, we work under the guidance of Dr. Soledad Funes, who is always ready to solve any doubts we have regarding the disposal of biological and non-biological wastes.

In our project is of outmost importance to consider the management and destination of the dangerous and toxic reagents, genetically modified organisms and other substances that may pose a threat to the environment and/or living beings. In the experiments we are performing, the presence of dangerous reagents is limited and therefore buffers can be easily disposed. As common practice, strong acids and bases are neutralized before disposal, and other reagents are disposed according to the manufacturer recommendations and to local laws and regulations. Regarding genetically modified organisms, we will be working with Escherichia coli DH5a that is not potentially pathogenic nor presents a health hazard neither to the researchers nor to the environment. According to GLP it is possible to handle this strain with the biosafety protocols of a basic laboratory (Level 1). Nevertheless, all the leftover media as well as the laboratory consumables used are autoclaved in order to prevent any uncontrolled propagation result of our work.

During the course of this project we always take safety measures in order to prevent any risk in the lab that may potentially lead to a hazardous accident. We believe that order and precaution are two basic principles for a good work in the laboratory: If one is able to foresee and prevent bad outcomes with good practices, a more efficient and safe environment can be achieved. This basic prevention measures can be, for example, the correct utilization of a fume hood when working with strong acids, bases and volatile substances; to handle cultures and media in a sterile atmosphere; the setup of an adequate space when working with a Bunsen burner; and the use of gloves, safety glasses and a lab coat in order to prevent exposure to chemicals.

iGEM Safety Key Questions

Biological sciences are a great tool for innovation and improvement of human society; nevertheless when the biosafety parameters are ignored, this area of research can become quite dangerous. We always have that idea present while working at the laboratory. We follow the rules stated at the Good Laboratory Practice (GLP) and according to the safety guidelines established by the Instituto de Fisiología Celular (IFC), part of the Universidad Nacional Autónoma de México (UNAM). We also have been working under the statutes of the Mexican official law (published in 2005) and guidelines (published in 2008 and reformed in 2009) about biosafety regarding genetically modified organisms, both available in the Mexican Government web site. Furthermore, we work under the guidance of Soledad Funes, PhD, who is always ready to solve any doubts we have regarding the disposal of biological and non-biological wastes.

The materials used throughout the project do not pose any safety or health risk neither to the team members nor to the rest of the members of the lab. For the development of this project, we have looked for those materials and reagents which would represent the least risk for ourselves, others and the environment. We understand, however, that when used in the wrong concentrations or without the proper care, some of the solutions, buffers and materials can be potentially harmful. In order to avoid any possible harm or injury, during every lab work session, all the team members always use gloves and white cotton laboratory coat as basic protection, and each time they start working or finish any experiment, common basic material, like micropipettes, UV transilluminator, analytical balance, used reagents, timers, glass ware and the lab bench, is cleaned superficially with water and/or ethanol 70%, to assure avoidance of any health risks to lab members as well to prevent cross-contamination among other experiments.

The materials and reagents used also do not represent any safety or health risk to the general public. It is important to stress that, like any other institutional research center, the access to the building is restricted and controlled, so general public entrance is not allowed, as well to children and suspect persons. With this measure, the possibility of accidents and malicious misuse of our experimental material by other individuals or groups is reduced.

Furthermore, they also do not pose any risk to the general public or to the environmental quality when released. During the development of our project, it is of outmost importance to consider the management and destination of the dangerous and toxic reagents, genetically modified organisms and other substances that may pose a threat to the environment and/or living beings. In the experiments we are performing, the presence of dangerous reagents is limited and therefore buffers and other non-biological reagents can be easily disposed. As common practice, strong acids and bases are neutralized before disposal, and other reagents are disposed according to the manufacturer recommendations and to local and institutional laws and regulations. All the used plastic material (like pipette tips, eppendorf tubes, etc.) is disposed into a special container and afterwards incinerated.

During the course of this project, we always take safety measures in order to prevent any risk in the lab that may potentially lead to a hazardous accident. We believe that organization and caution are two basic principles for a good work in the laboratory: If one is able to foresee and prevent bad outcomes with good practices, a more efficient and safe environment can be achieved, for the lab and outside the working space (environment). This basic prevention measures can be, for example, the correct utilization of a fume hood when working with strong acids, bases, volatile and flammable substances; to handle cultures and media in a sterile and controlled atmosphere; the setup of an adequate space when working with a Bunsen burner; and always prevent exposure to dangerous chemicals. Also, to prevent accidents related with all the equipment used in the project, we always use them strictly following the provider manuals, including centrifuges, gel cast for electrophoresis, incubators, etc.

None of the parts or devices used in this project are associated with, or known to cause pathogenicity, infectivity or toxicity, they also are not considered as a threat to environmental quality and they also do not pose any security concern. All our genetically modified organisms are hosted in the Escherichia coli DH5 strain, which is known to be a nonpathogenic bacterial strain widely used as a model organism. It is derived from the pathogenic bacteria E. coli, but this strain has been genetically modified to turn it not potentially pathogenic, so it isn’t a serious health risk, neither to the researchers, team members, lab member nor to the environment and society in case they are accidentally released. According the Good Laboratory Practices (GLP), it is possible to handle this strain with biosafety protocols of a basic laboratory (Level 1). Nevertheless, all the leftover media, as well as the discarded laboratory consumables, are disinfected with ethanol 70% immediately, disposed in chlorine for some hours, and finally autoclaved, in order to prevent any uncontrolled microbiological propagation derived from our work.

The IFC follows the biosafety regulations from the UNAM and in particular the recommendations made by the Environmental Management Unit (http://alexandria60.etcserver.com/~fqugau/residu2_archivos/ddeewsfcss/apoyo.html). A more detailed protocol for identification, evaluation, characterization and disposal of substances which might pose some kind of biological risk can be found here (http://www.biomedicas.unam.mx/_administracion/_unidades_apoyo_inst/manual_bioseguridad.pdf). These regulations include handling of the basic equipment in a laboratory, management and disposal of biological and chemical waste, correct behavior in the laboratory, how to behave in case of laboratory accidents, what to do in case of emergency (i.e. fires, earthquakes) and protection measures.

Before we have started our work in the lab, we were given an introductory lecture on the basic safety measurements we have to follow with special focus on the safety level we will be working (Biosafety Level 1). Some of the Mexican regulations on biosafety are summarized here (http://amexbio.org/links.html).

We think that biosafety must be always a central part of all projects, especially those of biological and chemical nature, because any research, besides their scientific or technological significance, can’t be allowed if it poses potential health risks and integrity threats to the lab team, general public or for environment. To reduce these risks, basic rules and safety guidelines, as stated previously in this document, as well as knowledge and training of who to act in case of accident or danger situations involving experimental material, is required and should be mandatory for institutions to provide to researchers, lab members, collaborators and students of biological and chemical sciences. With this, science and technology should achieve its goal of developing relevant knowledge and solutions, providing more benefits and fewer risks each time, in the way of improving life through knowledge and innovation.

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