Team:Bielefeld-Germany/Safety

From 2012.igem.org

Revision as of 19:53, 6 September 2012

1. Would any of your project ideas raise safety issues?

• researcher safety
• public safety
• researcher safety

the listet Organisms we use (or from which we use parts) are all part of the basic biosafety level 1:

• Pichia pastoris WT X33 ^[3]
• Pichia pastoris GS115 ^[3]
• Aequeora victoria ^[1]
• Cellulomonas fimi ATCC 484 ^[2]
• Clostridium cellulovorans ^[1]
• Escherichia coli BL21 (DE3)
• Thermus thermophilus HB27 ^[1]
• Bacillus halodurans C-125 ^[1]
• Bacillus pumilus DSM 27 (ATCC7061) ^[1]
• Xanthomonas campestris pv. campestris B100 ^[1]
• Streptomyces lavendulae REN-7
• Streptomyces griseus IFO 13350 ^[3]

For assuring the safety of the researcher every team member has to take part in special lab safety training. This training includes theoretical and practical parts where the team members learn how to handle (harmful) chemicals, rules for handling the setups and general safety regulations which have to be considered in the lab.

Further, everyone is required to be informed in detailed about the properties of every used chemical reagent. This implies that everyone has to read the R/S phrases and consider the recommended safety regulations.

To protect the researcher and the environment every person in the lab is required to wear safety clothes which implies long pants, closed shoes, a lab coat, safety glasses and when necessary gloves.

Other students and people without the authority and training do not have access to our iGEM lab. We take care that no materials which are biological modified leave the lab. In the case that samples, chemicals or something else need to be transferred to another room or even lab to continue an experiments we use special containers. For example, solvents are carried in baskets and samples in closed containers. Our project does not include the work with infectious organisms. Cell material can be thrown in the sink which includes a thermical disinfection system.

Even if the project goes according to plan we are exposed to harmful chemicals during the lab work. the most harmful chemicals are listed below with the material safety data sheet ([http://www.chemdb.de/ chemdb.de]):

• [http://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=DE&language=de&productNumber=46067&brand=SIGMA Ethidium bromide] for staining the DNA-gel
• [http://www.alfa-ebsoft.com/msds/pdf/german/L15075.PDF Acrylamide] in the SDS-PAGE
• [http://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=DE&language=de&productNumber=08253&brand=FLUKA Copper chloride] in the culture medium for the fermentation

To test our produced laccases we choses different substrates under which there are also some harmful compbounds:

• [http://de.wikipedia.org/wiki/Estradiol Estradiol]: toxic
• [http://de.wikipedia.org/wiki/Estron Estrone]: toxic
• [http://de.wikipedia.org/wiki/Ethinylestradiol Ethinylestradiol]: toxic
• [http://de.wikipedia.org/wiki/Diclofenac Diclofenac]: toxic
• [http://de.wikipedia.org/wiki/Ibuprofen Ibuprofen]: harmful to health
• [http://de.wikipedia.org/wiki/Naproxen Naproxen]: harmful to health
• [http://de.wikipedia.org/wiki/Naphthalin Naphthalene]: harmful to health and dangerous to environment
• [http://de.wikipedia.org/wiki/Acenaphthen Acenaphthene]: dangerous to environment
• [http://de.wikipedia.org/wiki/Phenanthren Phenanthrene]: dangerous to environment
• [http://de.wikipedia.org/wiki/Anthracen Anthracene]: irritant and dangerous to environment
• [http://de.wikipedia.org/wiki/Lindan Lindane]: toxic and dangerous to environment

These chemicals are stored in the lab regarding the safety regulations. Therefore, solvents are stored in explosion protected lockers and toxic chemicals are locked separately. Of course all chemicals are labeled adequate. Harmful chemicals, especially experiments which include the handling of solvents, are carried out under a fume hood. Experiments which require the protection from contaminations are carried out under a clean bench.

Some experiments are carried out in special areas in the lab or even in another room. One example is ethidium bromide which we mainly used for staining our gels. We only use the chemical in a special marked area of our lab and have separate equipment to prevent contaminations of other lab materials.

As already mentioned above, we also care about the environment. Therefore, all chemicals are disposed regarding their properties such as toxicity and explosiveness. All chemicals and chemically contaminated trash is disposed by the chemical disposal department. The department is also able to recycle solvents, such as ethanol and acetone (requires only a low grade of contamination). In this case we get the distilled solvents, labeled as technical solvents back in our lab and can use it for experiments or applications where the pureness is not a crucial factor.

If the project goes not according to plan there are following issues:

The laccases could be detached from the immobilization material for example if the CBD (cellulose binding domain) is not strong enough. When they are washed out in the drinking water there is the possibility that they will be consumed by the public. The use of laccases in the food industry ^[4] let us assume that there is no further issue of the consumption of laccases.

The laccases are known to have a broad spectrum of activity. The worst case is that the laccases will degrade estrogen or other chemical compbounds to toxic degradation products. To avoid this worst-case scenario the degradation products should be analysed before using it in the wastewater treatment plant.

In summary, we take care about the safety of ourselves, try to work as clean as possible and take care that the environment is protected as good as possible. The rules and safety measures we consider are standard at the Bielefeld University as in Germany.

References

[http://www.google.de/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0CCoQFjAA&url=http%3A%2F%2Fwww.opbw.org%2Fnat_imp%2Fleg_reg%2Fgerm%2FRegulations_Biol_Agents_bacteria.pdf&ei=jOY8UMasLarV4QThu4BQ&usg=AFQjCNFcxERwLdjdjNK8IdHyTs5a-FXjcg [1]] Einstufung von Bakterien in Risikogruppen, Bundesarbeitsblatt 10/2002

[http://www.cect.org/english/vbac.php?cect=4283&lan=en [2]] COLECCIÓN ESPAÑOLA DE CULTIVOS TIPO, www.cect.org/english/vbac.php?cect=4283&lan=en, on www.cect.org, Aug 30, 2012

[http://www.bvl.bund.de/SharedDocs/Downloads/06_Gentechnik/register_datenbanken/organismenliste.pdf;jsessionid=BBE29883867AB94FF1B68820F342520C.1_cid103?__blob=publicationFile&v=4 [3]] Bundesamt für Verbraucherschutz und Lebensmittelsicherheit, www.bvl.bund.de/DE/06_Gentechnik/03_Antragsteller/06_Institutionen_fuer_biologische_Sicherheit/01_ZKBS/03_Organismenliste/gentechnik_zkbs_organismenliste_node, on www.bvl.bund.de, Sep 5, 2012

[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2963825/ [4]] Johann F. Osma, José L. Toca-Herrera, Susana Rodriguez-Couto (2010), Uses of Laccases in the Food Industry, Enzyme Research 2010:1-8

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

• did you document these issues in the Registry?
• how did you manage to handle the safety issue?
• How could other teams learn from your experience?

No, not any of the new BioBrick parts that we made this year raise any safety issues. Although we are working with a plant pathogenous organism (Xanthomonas campestris pv. Campestris B100), but we are using a harmless enzyme laccase. Moreover the enzyme laccase of Trametes versicolor is intended for use in the food-processing industry, for example for bakery products and drinks. Furthermore the organisms we used to isolate laccases and our production strain Escherichia coli KRX are S1 organisms, which are not able to survive outside the lab conditions. This is a requirement for S1 organisms ([http://www.bvl.bund.de/SharedDocs/Downloads/06_Gentechnik/register_datenbanken/organismenliste.html?nn=1484542 list of these organisms]) defined by the german act of genetics ([http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 GenTSV]):

Biosecurity stage 1 Organisms

• are not human-, phyto- or animalpathogen
• do not contain or release organisms containing to a higher risk stage
• are approved by experiments or long term evaluation or do not proliferate in the enviroment because of biological implanted boundaries.

3. Is there a local biosafety group, committee, or review board at your institution?

• If yes, what does your local biosafety group think about your project?
• If no, which specific biosafety rules or guidelines do you have to consider in your country?

There is no Biosafety Committee at our university, only a safety officer. Nevertheless, we have an iGEM instructor, Dr. Jörn Kalinowski, who is kept informed about the project progress the entire time and is responsible for the biosafety of our project. Also, our university has a number of teams for waste management and fire safety.

The general rule at our university is that before every lab course every student has to take part in specific safety training for the course. Further, once a year, depending on the field of study and research, every member of the labs is required to take a refreshment course in lab safety, bio safety, fire safety, waste management, handling gas cylinders and working with hazard materials. All safety classes include a presentation and a practical part. For example in fire safety every student practices how to use the different types of fire distinguisher.

Before we (the iGEM team) started our lab work we took part at a general lab safety training course. Also, we had specific training for working with genetically modified organisms (GMOs) which is regulated in Germany by a number of laws. The work with GMOs has to be documented carefully. Handling, decontamination and disposal of GMOs is strictly regulated. According to the guidelines genetically modified organisms and contaminated materials can leave the lab after decontamination. We make sure that all of our work is done considering the relevant rules and regulations.

The [http://www.bvl.bund.de/DE/06_Gentechnik/gentechnik_node.html Federal office of consumer protection and safety] (Federal office of consumer protection and safety in [http://www.bvl.bund.de/EN/06_Genetic_Engineering/genetic_engineering_node.html English]) is the institution responsible for biosafety at universities, institutions and so on... All laws, regulations and guidelines can be found here:

• [http://www.bvl.bund.de/DE/06_Gentechnik/03_Antragsteller/08_Rechtsvorschriften/01_Deutschland/gentechnik_rechtsgrundlagen_national_node.html National German laws and regulations]
• [http://www.bvl.bund.de/DE/06_Gentechnik/03_Antragsteller/08_Rechtsvorschriften/02_Europa/gentechnik_rechtsgrundlagen_eu_node.html European regulations and guidelines]
• [http://www.bvl.bund.de/DE/06_Gentechnik/03_Antragsteller/08_Rechtsvorschriften/03_International/gentechnik_internationale_uebereinkuenfte_node.html International contracts and agreements]

The Federal office of consumer protection and safety appoints a team of experts ([http://www.bvl.bund.de/DE/06_Gentechnik/02_Verbraucher/05_Institutionen_fuer_biologische_Sicherheit/02_ZKBS/gentechnik_zkbs_node.html ZKBS], Zentrale Kommission für die Biologische Sicherheit) who determine the risks of GMOs for humans, animals and the environment. Based on their results they can suggest rules and regulations for handling GMOs.

ZKBS in [http://www.bvl.bund.de/EN/06_Genetic_Engineering/ZKBS/01_Allg_Stellungnahmen/genetic_engineering_zkbs_allgStellungnahmen_node.html English]

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

No, because the safety issues are well defined. It is really important, that every member get a Safety directive. Each of them should be fully aware and expressly assume the risks and danger in the laboratory work. Hazardous chemicals, like ethidium bromide are kept strictly separate from other chemicals (see Question 1).