Team:Evry/Safety

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Latest revision as of 21:45, 25 September 2012

We are working at the platform of Institute of Systems and Synthetic Biology (iSSB), which is a research unit of Université d’Evry-Val-d’Essonne. It is fully equipped for bacterial genetics and molecular biology work in terms of devices and has typical Biosafety Level 1 laboratories.

Our team is under daily supervision of Prof. Alfonso Jaramillo, the team direction of Bio-Synth Group of iSSB. We receive regular help as well from Prof. Ioana Popescu, engineer and hygiene and security correspondent, assisted by Dr. Dominique Zeliszewski, as well as from Thomas Landrain, a Ph.D. student working on site. All the iSSB research team is always around to help and advise us about good laboratory practices.

Safety page of the institute we are working in

http://wiki.issb.genopole.fr/index.php/HS:Main

http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf

Safety questions

1. Would any of your project ideas raise safety issues in terms of:

Researcher safety

Chassis

Escherichia coli: is the most commonly used gram-negative bacterial chassis in Molecular Biology. E. coli is commonly found in the lower intestine of warm-blooded organisms (endotherms). E. coli can benefit their hosts by producing Vitamin K2,and by preventing the establishment of other pathogenic bacteria within the intestine. We work with DH5a and TOP 10 that are common laboratory strains, considered as Level 1 Biosafety Containment agent and are non-pathogenic. However researchers are advised to use standard laboratory safety equipment and procedures while handling the cultures including wearing lab coats, gloves, etc.
Xenopus tropicalis: is a model organism for genetics, developmental biology, cell biology, toxicology, neuroscience and recently for synthetic biology. It is not dangerous for humans.

Dangerous substances

http://www.ozoneapplications.com/info/ozone_msds.jpg

All the standard molecular biology protocols we use in our experimentation are not dangerous for the researchers. However, our team works with potentially dangerous substances:

Ethidium Bromide (EtBr): is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) for agarose gel electrophoresis. Ethidium bromide may be a mutagen, carcinogen or teratogen although this depends on the organism and the conditions. EtBr intercalates double stranded DNA (i.e. inserts itself between the strands), deforming the DNA. This could affect DNA biological processes, like DNA replication and transcription.

Acute health effects: May cause eye and skin irritation. May cause gastrointestinal irritation with nausea, vomiting and diarrhea. May cause respiratory tract irritation. May cause methemoglobinemia, which is characterized by dizziness, drowsiness, headache, shortness of breath, cyanosis (bluish discoloration of skin due to deficient oxygenation of the blood), rapid heart rate and chocolate-brown blood.
Chronic health effects: May cause methemoglobinemia, which is characterized by chocolate-brown colored blood, headache, weakness, dizziness, breath shortness, cyanosis (bluish skin due to deficient oxygenation of blood), rapid heart rate, unconsciousness and possible death. May alter genetic material. Hence, we do not wash the gel in EtBr solution. Instead, EtBr is added in minimal concentration to the agar solution before it solidifies. It is strictly observed that gloves are used when handling EtBr or EtBr-containing gel. Gels, EtBr-containing gloves and tips are discarded considering them to be 'Hazardous Waste'.

Auxin extraction and identification methods such as HPLC, LC-MS/MS require using potentially dangerous substances. Lab coats, gloves are mandatory. Laminar hood is used when working with acetonitrile and methanol. Following there is a list of used chemical compounds for the mentioned methods and their potential health effects:

Acetonitrile: inflammable liquid.
- Acute effects: Hazardous in case of skin contact (irritant), of eye contact (irritant), of ingestion, of inhalation. Slightly hazardous in case of skin contact (permeator). Severe over-exposure can result in death.
- Chronic effects: The substance is toxic to blood, kidneys, lungs, liver, mucous membranes, gastrointestinal tract, upper respiratory tract, skin, eyes, central nervous system (CNS). The substance may be toxic to the reproductive system. Repeated or prolonged exposure to the substance can produce target organs damage. Repeated exposure to a highly toxic material may produce general deterioration of health by an accumulation in one or many human organs.
Methanol: flammable liquid and vapor. May be fatal or cause blindness if swallowed. Vapor harmful. Harmful if swallowed, inhaled, or absorbed through the skin. Causes eye, skin, and respiratory tract irritation. May cause central nervous system depression.
- Target Organs: Eyes, nervous system, optic nerve.
Formic acid: inflammable liquid, causes dangerous burns when get in eye- and skin contact.
- Acute effects: very hazardous in case of skin contact (irritant), of eye contact (irritant, corrosive), of ingestion. Hazardous in case of skin contact (corrosive, permeator). Slightly hazardous in case of inhalation (lung sensitizer). Non-corrosive for lungs. Liquid or spray mist may produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Inhalation of the spray mist may produce severe irritation of respiratory tract, characterized by coughing, choking, or shortness of breath. Inflammation of the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or, occasionally, blistering.
- Chronic effects: Slightly hazardous in case of skin contact (sensitizer). Mutagenic for mammalian somatic cells. Mutagenic for bacteria and/or yeast. The substance may be toxic to kidneys, liver, upper respiratory tract, skin, eyes, central nervous system (CNS). Repeated or prolonged exposure to the substance can produce target organs damage. Repeated or prolonged contact with spray mist may produce chronic eye irritation and severe skin irritation. Repeated or prolonged exposure to spray mist may produce respiratory tract irritation leading to frequent attacks of bronchial infection.
Perchloric acid: explosive liquid.
- Acute effects: very hazardous in case of skin contact (corrosive, irritant), of eye contact (irritant, corrosive), of ingestion,. Non-corrosive for lungs. Liquid or spray mist may produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Inhalation of the spray mist may produce severe irritation of respiratory tract, characterized by coughing, choking, or shortness of breath. Prolonged exposure may result in skin burns and ulcerations. Over-exposure by inhalation may cause respiratory irritation. Inflammation of the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or, occasionally, blistering.
Iron (III) chloride: corrosive. Harmful if swallowed. Causes eye and skin irritation and possible burns. May cause respiratory and digestive tract irritation. May cause blood abnormalities. May cause liver and kidney damage.
- Target Organs: Blood, kidneys, liver.
Sodium hydroxide: Harmful if swallowed. May cause adverse reproductive effects based upon animal studies. May cause liver and kidney damage. Causes burns by all exposure routes.
- Target Organs: Kidneys, liver, cardiovascular system. Causes eye and skin burns in contact, destructive to the tissue of the mucous membranes and airway, chronic exposition can cause lungs and ovaries cancer (tested on hamster).

http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf

Public safety

Any part of our project is not potentially dangerous or harmful for public. All DNA constuctions and E. coli expressing resistance genes (e.g. Ampicillin and Kanamycin) are disposed of properly, either by autoclaving or adding bleach.

Environmental safety

For our biological waste we are using the special plastic bags which are marked as biohazard, and which are disposed in the specially designated areas. All contaminated waste is treated as 'Bio-Hazardous Waste' and is decontaminated (autoclaved) before being put into a red trash bag or a sealed medical waste box. The bio-hazard bag is placed in a pre-chosen area for picking up. Appropriate measures for waste disposal are taken. Waste is not washed down the sink or thrown into public dust-bins. We are using embryos and tadpoles in laboratory conditions, they will never be released to the environment so we will not cause any environmental safety issues. They are going to be euthanized before reaching their reproductive age.

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, there are no potentially dangerous biobricks in our constructs. All DNA constuctions and E. coli expressing resistance genes are disposed of properly, either by autoclaving or adding bleach.

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?

In our host laboratory- iSSB there is no local committee which deals with the biosafety. However there is health and safety officer Prof. Ioana Popescu, Dr. Dominique Zeliszewski and people who are trained in first aid. Emergency numbers and first-aid rules are easily visible in each room of the institute. ISSB is a part of CNRS (French national center for research) .Consequently, the institute conforms to safety rules and regulations set by CNRS.
IGEM members have been biosafety-trained proior starting in vitro manipulations. More over, IGEM members have civil insurance in case of any lab accident. The permanent research team members are available throughout the time when experiments are conducted, and they offer supervision every now and then.
The laboratory has been classified as to biosafety categories and complies to French law (French Biosafety-National Legislation that deals with biohazards and GMO/GMM).
Environmental measures has been considered wastes are being disposed of in a specific containers and thereafter treated in accordance to the type of wastes whether biological or chemical.

http://nobel.scas.bcit.ca/debeck_pt/science/images/labsafety.jpg

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?

Safety in synthetic biology has always been both researchers’ and public’s concern. As a result, having an IGEM biosafety guidelines set for new IGEMers with what to use versus what is off limits. As for what can be done to make IGEM parts, devices, and systems safer, we propose the following potential solutions:

Introducing a CRISPR-induced system that targets and degrades plasmids and parts should it escape lab containment. The introduction of short sequences to all parts will trigger the CRISPR system if the antagonist is absent from the culture medium. This way, all parts and devices are restricted to laboratory containment.
Inserting killing genes (kill switch) as well as antibiotic resistant genes within all the constructs made for bacteria.
Designing a genetic switch mechanism making the frogs sterile, in case the tadpoles were issued to metamorphose into frogs, which could increase the risk of environmental dissemination.

@@ Line 1: / Line 1: @@
 {{:Team:Evry/template_v1}}
-Please get inspiration from this page to write our own safety page
+<html>
+<ul>
+We are following the official iGEM <a href="https://2012.igem.org/Safety">Safety instructions</a>.
-__NOTOC__
+<table>
+<tr>
+<td><p>We are working at the platform of <a href="http://www.issb.genopole.fr/">Institute of Systems and Synthetic Biology (iSSB)</a>, which is a research unit of Université d’Evry-Val-d’Essonne. It is fully equipped for bacterial genetics and molecular biology work in terms of devices and has <a href="http://www.who.int/csr/delibepidemics/WHO_CDS_CSR_LYO_2004_11/en/">typical Biosafety Level 1 laboratories</a>.
+<br/><br/>
+Our team is under daily supervision of Prof. Alfonso Jaramillo, the team direction of Bio-Synth Group of iSSB. We receive regular help as well from Prof. Ioana Popescu, engineer and hygiene and security correspondent, assisted by Dr. Dominique Zeliszewski, as well as from Thomas Landrain, a Ph.D. student working on site. All the iSSB research team is always around to help and advise us about good laboratory practices. </br></br>
+<b>Safety page of the institute we are working in</b></br>
+</p>
-=Safety=
+<a href="http://wiki.issb.genopole.fr/index.php/HS:Main">http://wiki.issb.genopole.fr/index.php/HS:Main</a></br></br>
-We are following the official iGEM [[Safety | safety instructions]] as well as the updated questions sent to us.<br>
+<td><img src="https://static.igem.org/mediawiki/2012/archive/3/30/20120907162311%21Lab1.png" style="width:350px">
-We are working in the [http://www.necker.fr/tamara/pages/people.html U1001 research unit] which is shared between iSSB (National Institute of Health and Medical Research) and [http://app.parisdescartes.fr/cgi-bin/WebObjects/Labs.woa/wa/showInfoLabo?cle=20101353#ficheLabo Paris Descartes University]. It is fully equipped for bacterial genetics and molecular biology work in terms of material and people having the competence to do the research. The lab is monitored on regular basis by security officers of both the Paris Descartes University and the iSSB.<br>
+<a href="http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf">http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf</a>
-Our team is under daily supervision of primarily Alfonso Jaramillo, Senior Research Scientist in U1001. We receive regular help as well from Aurore  (Engineer and Hygiene and Security Correspondent) as well from two Ph.D students Thomas Landrain and Antoine Decrulle, both working on site. The whole lab team is also here at times to explain and advise us.<br>
+</table>
-== Would the materials used in your project and/or your final product pose:==
+<h2>Safety questions </h2>
-===a. Risks to the safety and health of team members or others in the lab?===
+</br>
+<h3>1. Would any of your project ideas raise safety issues in terms of: </h3>
+</br>
+<h4>Researcher safety </h4>
+</br>
+<h5>Chassis </h5>
+We are working with level 1 safety organisms, which are not dangerous for researchers:
+<ul>
+<li> <i><b>Escherichia coli:</b></i> is the most commonly used gram-negative bacterial chassis in Molecular Biology. E. coli is commonly found in the lower intestine of warm-blooded organisms (endotherms). E. coli can benefit their hosts by producing Vitamin K2,and by preventing the establishment of other pathogenic bacteria within the intestine. We work with DH5a and TOP 10 that are common <a href="http://www.openwetware.org/wiki/E._coli_genotypes">laboratory strains</a>, considered as  <b>Level 1 Biosafety Containment agent</b> and are non-pathogenic. However researchers are advised to use standard laboratory safety equipment and procedures while handling the cultures including wearing lab coats, gloves, etc.
+<li><i><b>Xenopus tropicalis:</i></b> is a model organism for genetics, developmental biology, cell biology, toxicology, neuroscience and recently for synthetic biology. It is not dangerous for humans.
+</ul>
+</br>
+<h5>Dangerous substances </h5>
+<table align="right">
+<tr><td><img src="https://static.igem.org/mediawiki/2012/archive/7/72/20120907162827%21MSDS.png" width="300px" ></td></tr>
+<tr><td class="caption"><a href="http://www.ozoneapplications.com/info/ozone_msds.jpg">http://www.ozoneapplications.com/info/ozone_msds.jpg</a>
+</td></tr>
+</table>
-'''''Escherichia coli''''': we use DH5a and K12 which are common laboratory strains[http://www.openwetware.org/wiki/E._coli_genotypes], also considered as  '''Level 1 Biosafety Containment agent'''. <br>
-In our different sub-projects, we will not further exacerbate the already weak pathogenicity of ''Bacillus subtilis'': our different designs are aimed at proving the existence and caracterising the nanotubes. It ensues from fundamental research that was published earlier this year. It focuses on a natural phenonemon that ''B.subtilis'' seems to exhibit in period of stress.<br>
-To this end, we are going to use different antibiotic resistance as well as Green Fluroescent Protein variants to observe under the microscope the transfer of molecules. These two first designs use non conjugative plasmids. Albeit their resistance they do not pose any risks to the safety and health of the team/lab members since the constructs are under IPTG induction. <br>
-Other, mainly amplifiers in emitter-receptor systems or fluorescent proteins to monitor the expression of genes (See the [[Designs]] page) are used in the other sub-projects of characterization. Hence, we will not use or modify toxins (that ''B.subtilis'' does not naturally have [http://epa.gov/biotech_rule/pubs/fra/fra009.htm]) nor any potentially dangerous metabolites that could be present in this species. On the contrary, we aim at using external inducers such as hyperspank promoters (IPTG inducible) and orthogonal systems (such as T7 promoters) or even non natural systems (T7 amber suppressor tRNA) thus limiting risks to both the team and lab members .<br>
-In the very unlikely event (see section above) of an infection by either bacterium, a treatment with regular antibiotics is sufficient, antibiotics that are also natural.
-====References for risk assessment====
-# Farrar WE Jr. 1963. Serious Infections Due to “Non-Pathogenic” Organisms of the Genus ''Bacillus''. Am J Med. Vol:34. pp:134-41 [http://www.sciencedirect.com/science?_ob=MiamiImageURL&_imagekey=B6TDC-4BXGKV2-1B4-1&_cdi=5195&_user=658968&_pii=0002934363900470&_check=y&_origin=&_coverDate=01%2F31%2F1963&view=c&wchp=dGLzVlz-zSkzS&md5=5378af9f1b74b03445c94b3a24f99ac6&ie=/sdarticle.pdf PDF ]
-# Farrar WE , Reboli AC. 2006. The Genus ''Bacillus''-Medical. In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E, editors. Prokaryotes, Part 1, Section 1.2.  New-York: Springer. 609-630 [http://www.springerlink.com/content/mp23l6460306188q/fulltext.pdf PDF]
-# Turnbull, PCB. 1996. ''Bacillus''. In: Baron S, editor. Medical Microbiology. 4th edition Chapter 15. Galveston (TX): University of Texas Medical Branch at Galveston. [http://www.ncbi.nlm.nih.gov/books/NBK7699/]
-# ''Bacillus'' on Wikipedia [http://en.wikipedia.org/wiki/Bacillus]
-#Bacillus subtilis Final Risk Assessment [http://epa.gov/biotech_rule/pubs/fra/fra009.htm]
-# [http://www.openwetware.org/wiki/E._coli_genotypes E. coli genotypes]
-===b. Risks to the safety and health of the general public if released by design or accident?===
+<p>
-None of our designs have potential to harm the public if released by design or accident and even less since our lab is especially equipped for microbial manipulation and everything is done to avoid risk of release.<br>
+All the standard molecular biology protocols we use in our experimentation are not dangerous for the researchers. However, our team works with potentially dangerous substances:
-However, we are conscious that as any bacterium it can be hazardous. For instance, all traumatic wounds, infected burns and any serious lesions can potentially be a terrain for ''Bacillus'' strains but it is '''very''' rare.  When found, ''B.subtilis'' is usually mixed with other types of bacteria, it can cause a ''bacteremia'' but it is easily eradicated. <br>
+</p>
-'''Treatment''': ''subtilis'' like every soil  ''Bacillus'' species can be treated with non-ß-lactam antibiotics and ''Escherichia coli'' can be treated with regular antibiotics, which does not pose any problem to a doctor.<br>
-===c. Risks to environmental quality if released by design or accident?===
+<ul>
-None of our design or parts are hazardous to the environment since they mostly code for un-natural systems or detection molecules. If anything they would have a disadvantage for overproducing useless metabolites such as GFP, their competitiveness is decreased. Any kind of horizontal transfer risks is taken into acount with the use of non-conjugative plasmids and under external inducers.
+  <li> <b>Ethidium Bromide (EtBr):</b> is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) for agarose gel electrophoresis.  Ethidium bromide may be a mutagen, carcinogen or teratogen although this depends on the organism and the conditions.  EtBr intercalates double stranded DNA (i.e. inserts itself between the strands), deforming the DNA. This could affect DNA biological processes, like DNA replication and transcription.</li>
+    <ul>
+      <li><b>Acute health effects:</b></i> May cause eye and skin irritation. May cause gastrointestinal irritation with nausea, vomiting and diarrhea. May cause respiratory tract irritation. May cause methemoglobinemia, which is characterized by dizziness, drowsiness, headache, shortness of breath, cyanosis (bluish discoloration of skin due to deficient oxygenation of the blood), rapid heart rate and chocolate-brown blood.</li>
-===d. Risks to security through malicious misuse by individuals, groups or states?===
+      <li><b>Chronic health effects:</b></i> May cause methemoglobinemia, which is characterized by chocolate-brown colored blood, headache, weakness, dizziness, breath shortness, cyanosis (bluish skin due to deficient oxygenation of blood), rapid heart rate, unconsciousness and possible death. May alter genetic material. Hence, we do not wash the gel in EtBr solution. Instead, EtBr is added in minimal concentration to the agar solution before it solidifies. It is strictly observed that gloves are used when handling EtBr or EtBr-containing gel. Gels, EtBr-containing gloves and tips are discarded considering them to be 'Hazardous Waste'.</li>
-Biosecurity is respected since no participants to the Paris 2011 Bettencourt team has any conflict of interests of any kind. In addition, there could not be security concerns due to a malicious use of our designs and parts/devices since they only aim at visualising a natural phenomenon present in ''B.subtilis'', it remains fundamental research with no potential misuse outside the laboratory. <br>
+    </ul>
-----
+  </li>
+</ul>
-===Final Statement on Biosafety and Biosecurity===
+<br/>
-<br> '''<big>In conclusion, not a single part nor a single system/final product is known to be infective/toxic/pathogenic to humans or threatening to the environment; there would be hence <u>no implications in case of release</u>.</big>''' <br>
-== Under what biosafety provisions will / do you operate?==
-===a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible. ===
-Our institution, iSSB, provides to its researchers, engineers and technicians a set of of regulations, advice and mandatory practices for each aspects of the laboratory life.
-Here is the link to all the online documentation: <br>
-http://www.rh.iSSB.fr/iSSB/IntraRh/RHPublication.nsf/mDisplayMotsClefsWeb?OpenForm&arg1=6&arg2#
-<br>
-You can find the relevant information in the tab "Formation - Documentation". Our apologies for the site being in French but you can have some of the safety and advice sheets in English by clicking [http://www.rh.iSSB.fr/iSSB/IntraRH/RHPublication.nsf/vMotsClefsWebByCat/0735F4BC31ADA4BEC125756A005246A2/$file/Anglais.zip?OpenElement on this].
-In essence, there are information and rules concerning cleanliness and important reflexes to have; it treats on how to sterilize and wash contaminated material. There is also a part concerning hazardous situations (electrical, chemical, ionizing, fire etc.) and how to deal with them, who to contact, how to deal with dangerous situations as well as your rights and obligations such as the one to refuse to enter your workplace if there is potential threat to your place and the obligation to contact the director and the Head Safety Officer of the iSSB that will emit the final judging. A good point is always to tell your hierarchy superior and the lab security officer.
-===b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review. ===
-The iGEM team as a project, is part of the four-year work plan of our hosting U1001 iSSB lab that was reviewed for its scientific merit and safety of the research plan. There was no specific contact with iSSB/University institutional biosafety personnel regarding our specific project that was reviewed by the lab heads and accepted by the lab safety officer - it is within his responsibilities as the representative of the institution to provide such agreement after reviewing our project.
-===c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.===
-We received a laboratory training '''prior''' to the start of the experiments.<br>
-The information about biosafety in the lab presented here have been gathered during the introduction made by the Correspondant of Hygiene and Security ([http://www.necker.fr/irnem/units/search.php?iduser=1950&pass=1 Aurore ]) to the safe use of each instrument and machine as well as general guidelines of good lab practice. All the students in the team participated to this introduction. The handling of waste, how to keep things sterile, how to use the hood etc. was also explained to us.<br>In our team, there are also two designated ''lab managers'' that are notably in charge of knowing the functioning of each apparatus and of the security issues; team's students had, as a mandatory part of their undergraduate curriculum, a Hygiene and Security course. In case any problems arise Aurore , the lab's security officer,  can be contacted 24H/24H to seek help and/or advice. Furthermore, weekly team meetings with our supervisors allow for any concerns to be raised and treated.
-===d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.===
-France, as a member of the European Union follows the Directives of the European Parliament (such as this one [http://www.bmwf.gv.at/fileadmin/user_upload/forschung/gentechnik/2009-41-EC.pdf Directive 2009/41/EC on the contained use of genetically modified micro-organisms]). We, and the research unit we work in, apply the general guidelines that are outlined notably in the aforementioned Directive notably in terms of waste management and sterility. <br>
-==Safety at work==
-There are basic precautions and security procedures that '''MUST''' be followed in any laboratory and here are some that we gathered from this introduction and from later meetings with the security officer. <br>
-*'''The ten general rules :'''
- Rule n°1: If you're not sure of anything: ASK !
- Rule n°2 : Don't smoke or eat or drink in the lab.
- Rule n°3 : Don't wear scarves and tie your hair back if needed. Wear coat when it’s not so hot.
- Rule n°4 : Don't leave any personal belongs (phone...) because of thiefs.
- Rule n°5 : Don't work ALONE at night and during the weekend (and inform the on-call guard of your presence).
- Rule n°6 : Be careful, products may be harmful even if they seem inoffensive.
- Rule n°7 : Identify your stuff (iGEM + date + if sterile) and CLEAN UP once you've finished.
- Rule n°8 : Tell us if something is running out so that we can buy them in advance.
- Rule n°9 : Close both lab doors before leaving.
- Final rule : Turn off all the machines before leaving (especially the bain marie,
- because of fire and surtension problems in this lab!). Check that -20°C and -80°C are OK every morning.
-If you've received chemical spatter on your eyes or your skin, use ONLY water to clean them (at least 10 min under the water)! No detergent!<br>
-*'''Trash can :'''
-''Solid carton:'' Press down and compress! <br>
-Black: For everyday use  – paper, plastic, enveloppe...<br>
-Cardboard yellow box: Biological stuff (Petri dish closed, pipette on verticle angle)<br>
-To close the box: Read the instructions on the box, identify it (U1001 + date), then put it in the hallway<br>
-Plastic yellow box: Sharp/ cutting items (needles, glass...)<br>
-''Liquid :''
-Methanol and other: chemical waste bottle<br>
-*'''BET bench :'''
-ASK Aurore before using - Cancer Risk (UV and BET)!<br>
-Wear nitrile blue or purple glove, long-sleeved blouse and goggles<br>
-No BET in electrophoresis, only in tray.<br>
-Use only dishes, pipettes and trash can for BET. <br>
-Buffer : 0,5X (be careful if 1X = crystal) TBE (or TAE)
-''Specials trashes for EtB:''
-Solid (gel, gloves, contamined stuff) in big white box (under the bench)<br>
-Tips in yellow box on the bench “EtB”<br>
-Liquid: in 5L plastic bottle on the bench (decontamination with 2 bags of charcoal)<br>
-*'''Lavery :'''
-In Dirty stuff  Box (Buffer …): Rinse bottle and take out scotch<br>
-In Contaminated stuff  Box (bacteria …): Let it in the box. (It will be wash with Javel water)<br>
-If you decontamined with Javel water, wear blouse.<br>
-*'''Microscopy :'''
-ASK MING for a briefing before using! (his holidays: 2-17 of July) <br>
-Verify schedule before entry in the room. Do not let the door open (night for experiments which are on, air conditioned room)<br>
-*'''Machines instructions :'''<br>
-'''Autoclave :'''<br>
-To be used only by COMPETENT AND AUTHORIZED PEOPLE : Kevin and Daniel < risk of burning and glass breaking.<br>
-Always UNSCREW the bottles before introducing them into the machine! <br>
-Do not try to open during each cycle. Check  temperature (85°C) and  pressure of autoclave (1 bar) before opening.<br>
-Be careful, the top is very hot (high risk of burning yourself, 85°C when opened): Use mittens to take out the sterilized bottles! Check if the scotch lines turned black.<br>
-If it's LBA bottle, mix it after sterilizing.
-'''Micro-waves :'''<br>
-Always UNSCREW the bottles before introducing them into the machine! <br>
-BIG RISK OF BURNING: Use mittens to take out the bottles!
-'''Washing machine :'''<br>
-To be used only by competant and authorized people (Kevin and Daniel) < risk of burning and glass breaking.<br>
-Check the level of water before use (the reserve as to be full). Do not try to open during each cycle.
-'''Osmosed water recipient :'''<br>
-Be careful to close it well (90°) to avoid flood.
-'''Precision balance :'''<br>
-Prepare what you need and close doors everytime before weighing. Clean after use.
-'''Shaker-Incubator :'''<br>
-Rack has to be stable (check that black screws are on tight)<br>
-Don't hesitate to launch a new one if necessary.<br>
-Sticky band : don't put your hand on it (there is polystyrene stuff on it). To wash it, use only water + soap.<br>
-Always incline your falcon and unscrew it a little (to let the air go inside)
-'''37°C and 30°C Incubators :'''<br>
-Be careful when closing the door, it is fragile !<br>
-Petri dishes with gelose has to be put on the top (to avoid deshydration)
-'''Hood :'''<br>
-It only protects the area under the hood and not us ! NO PATHOGEN INSIDE!<br>
-Clean your hands. Clean the work area with 70% ethanol BEFORE and AFTER use.<br>
-(ethanol in 5L bottle (kitchen) + osmosed water)<br>
-Try to occupate only  middle of area in order to share with someone else.
-'''Centrifuges :'''<br>
-Don't forget to put the hat and to BALANCE out your tubes (weight and symmetric position).<br>
-Once you've finished, shut the machine down. Leave the top open.
-'''Freezers :'''<br>
-Use the -80°C only for long term stocks. Use -20°C for working stocks.<br>
-Don't leave the door open when you looking for a strain in the boxes. <br>
-Close the door of the “kitchen”: this room is air conditioned because of the -80°C!<br>
-Use cardboard boxes for storage (not plastic racks).<br>
-Always check the temperature and if there is a problem of cut, do NOT open the door (years of research are inside...)<br>
-If there is any problem: 2255 (technical service of the faculty:  you will join Jean Marie / Florent electricians or Eric (boss)).<br>
-°C - Powder of IPTG, Antibiotics … : Use glove
-'''Burner :'''<br>
-Sterilizes an area of 30 cm radius around the burner. Don't speak when working within the sterile zone, otherwise you'll spread your bacteria !<br>
-Be careful at your hear
-'''Bain Marie :'''<br>
-DO NOT FORGET to turn it off once you're done (high risk of combustion!)
-'''PCR / Thermocycler :'''<br>
-Don't let it work at 4°C overnight as much as possible (electricity surtension)! Shut it down after use.
-'''Biophotometer :'''<br>
-/10 dilution in 50 microL. Do not forget to shut down and close the chamber with the black cap.
-'''Spectrophotometer and Tecan :'''<br>
-Turn them off  before leaving.
-<html>
-<!-- PAGE FOOTER -- ITEMS FROM COLUMN ! HAVE BEEN MOVED HERE  -- RDR  -->
-        <div id="footer-wrapper">
+<p>
-        <div id="footer">
+Auxin extraction and identification methods such as HPLC, LC-MS/MS require using potentially dangerous substances. Lab coats, gloves are mandatory. Laminar hood is used when working with acetonitrile and methanol. Following there is a list of used chemical compounds for the mentioned methods and their potential health effects:
-              <div id="f-poweredbyico"><a href="http://www.mediawiki.org/"><img src="/wiki/skins/common/images/poweredby_mediawiki_88x31.png" height="31" width="88" alt="Powered by MediaWiki" /></a></div> 	      <div id="f-copyrightico"><a href="http://creativecommons.org/licenses/by/3.0/"><img src="http://i.creativecommons.org/l/by/3.0/88x31.png" alt="Attribution 3.0 Unported" width="88" height="31" /></a></div> 	     <ul id="f-list">
+</p>
+<ul>
+  <li> <b>Acetonitrile:</b> inflammable liquid.
+    <ul>
+      <li><b>Acute effects:</b> Hazardous in case of skin contact (irritant), of eye contact (irritant), of ingestion, of inhalation. Slightly hazardous in case of skin contact (permeator). Severe over-exposure can result in death.</li>
+      <li><b>Chronic effects:</b> The substance is toxic to blood, kidneys, lungs, liver, mucous membranes, gastrointestinal tract, upper respiratory tract, skin, eyes, central nervous system (CNS). The substance may be toxic to the reproductive system. Repeated or prolonged exposure to the substance can produce target organs damage. Repeated exposure to a highly toxic material may produce general deterioration of health by an accumulation in one or many human organs.</li>
+    </ul>
+  </li>
-	   <!-- Recentchanges is not handles well DEBUG -->
+  <li> <b>Methanol:</b> flammable liquid and vapor. May be fatal or cause blindness if swallowed. Vapor harmful. Harmful if swallowed, inhaled, or absorbed through the skin. Causes eye, skin, and respiratory tract irritation. May cause central nervous system depression.
-		     <li id="t-recentchanges"><a href="/Special:RecentChanges"
+    <ul>
-		      title='Recent changes'>Recent changes</a></li>
+      <li><b>Target Organs:</b> Eyes, nervous system, optic nerve.</li>
+    </ul>
+  </li>
+  <li> <b>Formic acid: </b>inflammable liquid, causes dangerous burns when get in eye- and skin contact.
+    <ul>
+      <li><b>Acute effects:</b> very hazardous in case of skin contact (irritant), of eye contact (irritant, corrosive), of ingestion. Hazardous in case of skin contact (corrosive, permeator). Slightly hazardous in case of inhalation (lung sensitizer). Non-corrosive for lungs. Liquid or spray mist may produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Inhalation of the spray mist may produce severe irritation of respiratory tract, characterized by coughing, choking, or shortness of breath. Inflammation of the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or, occasionally, blistering.</li>
-		     <li id="t-whatlinkshere"><a href="/Special:WhatLinksHere/Team:Paris_Bettencourt/Modeling"
+      <li><b>Chronic effects:</b> Slightly hazardous in case of skin contact (sensitizer). Mutagenic for mammalian somatic cells. Mutagenic for bacteria and/or yeast. The substance may be toxic to kidneys, liver, upper respiratory tract, skin, eyes, central nervous system (CNS). Repeated or prolonged exposure to the substance can produce target organs damage. Repeated or prolonged contact with spray mist may produce chronic eye irritation and severe skin irritation. Repeated or prolonged exposure to spray mist may produce respiratory tract irritation leading to frequent attacks of bronchial infection.</li>
-		      title="List of all wiki pages that link here [j]" accesskey="j">What links here</a></li>
+    </ul>
+ </li>
+ <li> <b> Perchloric acid: </b> explosive liquid.
+    <ul>
+      <li><b>Acute effects: </b> very hazardous in case of skin contact (corrosive, irritant), of eye contact (irritant, corrosive), of ingestion,. Non-corrosive for lungs. Liquid or spray mist may produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Inhalation of the spray mist may produce severe irritation of respiratory tract, characterized by coughing, choking, or shortness of breath. Prolonged exposure may result in skin burns and ulcerations. Over-exposure by inhalation may cause respiratory irritation. Inflammation of the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or, occasionally, blistering.</li>
+    </ul>
+  </li>
+  <li> <b> Iron (III) chloride: </b> corrosive. Harmful if swallowed. Causes eye and skin irritation and possible burns. May cause respiratory and digestive tract irritation. May cause blood abnormalities. May cause liver and kidney damage.
+    <ul>
+      <li><b> Target Organs:</b> Blood, kidneys, liver.</li>
+    </ul>
+  </li>
+  <li> <b>Sodium hydroxide: </b> Harmful if swallowed. May cause adverse reproductive effects based upon animal studies. May cause liver and kidney damage. Causes burns by all exposure routes.
+    <ul>
-                         <li id="t-recentchangeslinked"><a href="/Special:RecentChangesLinked/Team:Paris_Bettencourt/Modeling"
+      <li><b>Target Organs: </b></i>Kidneys, liver, cardiovascular system. Causes eye and skin burns in contact, destructive to the tissue of the mucous membranes and airway, chronic exposition can cause lungs and ovaries cancer (tested on hamster).</li>
-                          title="Recent changes in pages linked from this page [k]" accesskey="k">Related changes</a></li>
+    </ul>
+  </li>
+</ul>
+<br/></br>
+<table align="right">
+<tr><td><img src="https://static.igem.org/mediawiki/2012/b/b3/Biohazard.png" width="380px" align="right"></td></tr>
+<tr><td class="caption"><a href="http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf">http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf</a>
+</td></tr>
+</table>
+<h4>Public safety </h4>
+</br>
+<p>
+Any part of our project is not potentially dangerous or harmful for public. All DNA constuctions and <i>E. coli</i> expressing resistance genes (e.g. Ampicillin and Kanamycin) are disposed of properly, either by autoclaving or adding bleach.
+</p>
-	                 <li id="t-upload"><a href="/Special:Upload"
+<br/>
-	                  title="Upload files [u]" accesskey="u">Upload file</a>
+<br/>
-	                 </li>
+<h4>Environmental safety </h4>
-	                 <li id="t-specialpages"><a href="/Special:SpecialPages"
+</br>
-	                  title="List of all special pages [q]" accesskey="q">Special pages</a>
+<p>For our biological waste we are using the special plastic bags which are marked as biohazard, and which are disposed in the specially designated areas. All contaminated waste is treated as <b>'Bio-Hazardous Waste'</b> and is decontaminated (autoclaved) before being put into a red trash bag or a sealed medical waste box. The bio-hazard bag is placed in a pre-chosen area for picking up. Appropriate measures for waste disposal are taken. Waste is not washed down the sink or thrown into public dust-bins.
+We are using embryos and tadpoles in laboratory conditions, they will never be released to the environment so we will not cause any environmental safety issues. They are going to be euthanized before reaching their reproductive age.</p>
-	                 </li>
+<br/></br>
-                 <li><a href='/Special:Preferences'>My preferences</a></li>
+<h3>2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? </h3>
-             </ul>
+<h4>If yes,
-        </div> <!-- close footer -->
+did you document these issues in the Registry?
-        <div id='footer'>
+how did you manage to handle the safety issue?
-	     <ul id="f-list">
+How could other teams learn from your experience?</h4>
+<br/>
+<p>
+No, there are no potentially dangerous biobricks in our constructs. All DNA constuctions and <i>E. coli</i> expressing resistance genes are disposed of properly, either by autoclaving or adding bleach.
+</p>
+<br/></br>
+<h3>3. Is there a local biosafety group, committee, or review board at your institution? </h3>
+<h4>If yes, what does your local biosafety group think about your project?</h4>
+<h4>If no, which specific biosafety rules or guidelines do you have to consider in your country?</h4>
+<br/>
+<p>
+In our host laboratory- iSSB there is no local committee which deals with the biosafety. However there is health and safety officer Prof. Ioana Popescu, Dr. Dominique Zeliszewski and people who are trained in first aid. Emergency numbers and first-aid rules are easily visible in each room of the institute. ISSB is a part of CNRS (French national center for research) .Consequently, the institute conforms to safety rules and regulations set by CNRS. </br>
-	             <li id="t-print"><a href="/wiki/index.php?title=Team:Paris_Bettencourt/Modeling&amp;printable=yes"
+IGEM members have been biosafety-trained proior starting <i>in vitro</i> manipulations. More over, IGEM members  have civil  insurance in case of any lab accident. The permanent research team members are available throughout the time when experiments are conducted, and they offer supervision every now and then. </br>
-	              title="Printable version of this page [p]" accesskey="p">Printable version</a>
-	             </li>
+The laboratory has been classified as to biosafety categories and complies to French law (French Biosafety-National Legislation that deals with biohazards and GMO/GMM). </br>
+Environmental measures has been considered  wastes are being disposed of in a specific containers and thereafter treated in accordance to the type of wastes whether biological or chemical.
-	             <li id="t-permalink"><a href="/wiki/index.php?title=Team:Paris_Bettencourt/Modeling&amp;oldid=86565"
+<br/></br>
-	              title="Permanent link to this revision of the page">Permanent link</a>
+<table align="left">
-	             </li>
+<tr><td><img src="https://static.igem.org/mediawiki/2012/1/18/SafetyImg.jpg" width="340px" ></td></tr>
+<tr><td class="caption"><a href="http://nobel.scas.bcit.ca/debeck_pt/science/images/labsafety.jpg">http://nobel.scas.bcit.ca/debeck_pt/science/images/labsafety.jpg</a>
+</td></tr>
+</table>
+</p>
+<h3>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? </h3>
+<br/>
+<p>
+Safety in synthetic biology has always been both researchers’ and public’s concern. As a result, having an IGEM biosafety guidelines set for new IGEMers with what to use versus what is off limits. As for what can be done to make IGEM parts, devices, and systems safer, we propose the following potential solutions:
+</p>
- 		        <li id="privacy"><a href="/2011.igem.org:Privacy_policy" title="2011.igem.org:Privacy policy">Privacy policy</a></li>
+<ul>
-		        <li id="disclaimer"><a href="/2011.igem.org:General_disclaimer" title="2011.igem.org:General disclaimer">Disclaimers</a></li>
+<li> Introducing a CRISPR-induced system that targets and degrades plasmids and parts should it escape lab containment. The introduction of short sequences to all parts will trigger the CRISPR system if the antagonist is absent from the culture medium. This way, all parts and devices are restricted to laboratory containment.</li>
-	     </ul>
+<li> Inserting killing genes (kill switch) as well as antibiotic resistant genes within all the constructs made for bacteria.</li>
+<li> Designing a genetic switch mechanism making the frogs sterile, in case the tadpoles were issued to metamorphose into frogs, which could increase the risk of environmental dissemination.</li>
+</ul>
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+<script type="text/javascript">writeFooter()</script>
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