Team:UFAM Brazil/Safety

From 2014.igem.org

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<p><b>Sanitary requirements for the handling of mercury in laboratories (Applied to health labs).</b></p>
<p><b>Sanitary requirements for the handling of mercury in laboratories (Applied to health labs).</b></p>
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<p>Masks , Gloves , Lab coat.</p>
<p>Masks , Gloves , Lab coat.</p>
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<p>Use masks and goggles to protect eyes, mouth and nose against volatile chemicals, splashes and inhalation of aerosol particles. Use an autoclave to make sterile infectious material by effective heat. Use of bottles with screw cap to prevent spills and aerosols. Countertops are decontaminated before and after the development of some activities. Before developing the experiment, UV light cabin remains on for 15 minutes. Alcohol and detergent are used as chemical agents for cleaning hands against microorganisms.</p>
<p>Use masks and goggles to protect eyes, mouth and nose against volatile chemicals, splashes and inhalation of aerosol particles. Use an autoclave to make sterile infectious material by effective heat. Use of bottles with screw cap to prevent spills and aerosols. Countertops are decontaminated before and after the development of some activities. Before developing the experiment, UV light cabin remains on for 15 minutes. Alcohol and detergent are used as chemical agents for cleaning hands against microorganisms.</p>
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<p>Floor and Bench</p>
<p>Floor and Bench</p>
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<p>The floor of the room should be waterproof, without pores and cracks and easily cleanable. The work bench smooth and easy to clean.</p>
<p>The floor of the room should be waterproof, without pores and cracks and easily cleanable. The work bench smooth and easy to clean.</p>
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<p>The storage of mercury should be in unbreakable and airtight jars.  
<p>The storage of mercury should be in unbreakable and airtight jars.  
Code ADR: 8,66 To carry the mercury in a liquid state. Stored mercury must be labeled with the phrases: R23 (" Toxic by inhalation") and R33 ("Danger of cumulative effects").  
Code ADR: 8,66 To carry the mercury in a liquid state. Stored mercury must be labeled with the phrases: R23 (" Toxic by inhalation") and R33 ("Danger of cumulative effects").  
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<p>Disposal of waste mercury is considered biohazardous. Therefore, they must be placed in unbreakable bottles containing water inside and are sealed, labeled "biohazardous" and placed in a plastic bag milky white.
<p>Disposal of waste mercury is considered biohazardous. Therefore, they must be placed in unbreakable bottles containing water inside and are sealed, labeled "biohazardous" and placed in a plastic bag milky white.
All generated waste, such as cultures and stocks, microorganism culture media, the instruments used to inoculate cultures and biological samples are placed in order to be treated by a physical process to reduce or eliminate the microbial load. The sharps are discarded in rigid containers resistant to puncture, break and leak and properly autoclaved before disposal.
All generated waste, such as cultures and stocks, microorganism culture media, the instruments used to inoculate cultures and biological samples are placed in order to be treated by a physical process to reduce or eliminate the microbial load. The sharps are discarded in rigid containers resistant to puncture, break and leak and properly autoclaved before disposal.
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<p>Species name</p>
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</td>
<td width="20%" align="center">
<td width="20%" align="center">
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<p>PPE</p>
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<p>Risk Group</p>
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<p>Masks , Gloves , Lab coat.</p>
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<p>How it was acquired </p>
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<td width="45%"  align="justify" style="padding:10px">
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<p>Disease risk to human</p>
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<p>E coli RR1</p>
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<td width="20%" align="center">
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<p>1 <a href="http://www.absa.org/index.html" target="_blank">(Link)<a></p>
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<td width="20%" align="center">
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<p>genes were synthesized</p>
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<td width="45%"  align="justify">
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<p>Highly unlikely to cause disease in healthy laboratory workers, animals. But may cause irritation to skin, eyes, and respiratory tract if the contact is direct</p>
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<td width="15%" align="center">
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<p>E coli DH10B</p>
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</td>
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<td width="20%" align="center">
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<p>1 <a href="http://www.absa.org/riskgroups/bacteriasearch.php?genus=Escherichia" target="_blank">(Link)<a></p>
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<p>Assembly</p>
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<td width="45%"  align="justify" style="padding:10px">
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<p>May cause irritation to skin, eyes, and respiratory tract</p>
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<p>E coli DH5 alpha</p>
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<td width="20%" align="center">
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<p>1 <a href="http://www.absa.org/riskgroups/bacteriasearch.php?genus=Escherichia" target="_blank">(Link)<a></p>
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</td>
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<td width="20%" align="center">
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<p>genes were synthesized</p>
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</td>
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<td width="45%"  align="justify" style="padding:10px">
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<p>May cause irritation to skin, eyes, and respiratory tract</p>
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</td>
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</tr>
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<tr>
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<td width="15%" align="center">
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<p>E coli JM110</p>
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</td>
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<td width="20%" align="center">
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<p>1 <a href="http://ors.uchc.edu/bio/resources/pdf/3.6.1.A_colipath.pdf" target="_blank">(Link)<a></p>
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</td>
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<td width="20%" align="center">
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<p>genes were synthesized + BBa_E0840 or BBa_K346004</p>
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<td width="45%"  align="justify" style="padding:10px">
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<p>Present no hazard to human health, non-pathogenic, non-toxicogenic. Cause irritation to skin, eyes, and respiratory tract if the contact is direct</p>
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Latest revision as of 18:37, 10 October 2014

Biosafety

Mercury disposal in the environment

Currently, Brazilian legislation is regarding to the levels of metals released into the environment, but not to the laws on the handling of toxic metals such as Mercury. Besides the error in the use of mercury in terms of quantity, there is another contradiction of laws related to the regulation since the national legislation which governs the drinking water is the Ministry of Health and the one to rule over the aquatic environmental systems is the National Environmental Council (CONAMA).

Based on CONAMA N° 357, March 17th, 2005, the classification of waters is valid for the entire national territory. The classification is done in 9 classes, and the resolution separates fresh water (salinity < 0.05 %) in five types: Special Class and Class 1 to 4, in which the restrictions impurities decrease as the number of the class increases. Saline waters are classified as Class 5 and 6, and brackish waters are Class 7 and 8. The resolution also determines the maximum allowable levels for effluent discharge into the waters, except for special class of waters that cannot receive releases wastewater as it is intended for domestic supply and should be absent of coliforms. For Class 1, the maximum mercury content allowed is 0.2 mg/L for Class 2 is 2 mg/L, for Classes 5 to 7 is 0.1 mg/L, and for other classes there’s no maximum levels specified.

For other waterbody (class 1-8), which are neither the sub-surface nor special class, is tolerated dumps that meet the specifications of maximum (Hg = 0.01 mg/L) and has a maximum flow of 1.5 times of the activity average daily period of agent launcher, as soon as they do not exceed the limits set for their classes. Thus, legislation allows a higher concentration in the effluent than in the waterbody, believing that there is dilution of which was released. However, it is known that there are many sources of pollution that contains the same harmful substance that discharge their effluent into the same body of water, this fact can lead to concentration of substance and exceed limits of resolution.

In summary, Brazilian legislation only attempts to control the discharge of effluents, it does not act in maintaining the environmental quality of the receiving waterbody. Another mistake made by the legislation is that it rules on concentration and not on the load of harmful substance, this makes any dump mercury is disposed within the parameters in a greater amount than another source that turns your mercury with a margin slightly above established standards. Thus, whoever offers most eviction and degrades the bodies of water cannot be penalized, while those who dumps acceptable amounts will be.

Mercury disposal in the lab environment

Mercury is a silver metal that is liquid at normal temperature and odorless, when the temperature is increased becomes more dense toxic and corrosive. It is a product harmful when inhaled, ingested or in contact, causing irritation to skin, eyes and respiratory system. The mercury is present in various forms (metallic, organic, inorganic Hg) and can be found in three oxidation states (0, +1 , +2) generally readily interconvertible in nature.

Both humans and animals are exposed to all forms through the environment. The mercury vapor present in the atmosphere is eventually converted to water soluble form and returns to the earth's surface in rainwater. At this point, two important chemical changes may occur. The metal can be reduced again to mercury vapor and returns to the atmosphere or can be methylated by microorganisms in sediment from water, including fresh or sea water.

The handling of heavy metals cannot be made in ordinary laboratories, without any degree of certainty (See Table below about the requirements). In the absence of a suitable laboratory handling heavy materials at the university, the team had to redesign one of UFAM_Brazil laboratories (read more) to use mercury. This lab will remain for future research conducted at the university for now on.

Safety in the tests:

To avoid harmful we have used mercury in a water solution. In its ionic form Hg2+ in Mercury(II) Chloride form (HgCl2), though stock solution to test our biobricks. To do it, we will only have to dilute the mercury to the wanted concentration using sterilized water. The disposal have been done following the sanitary rules.

What the law says?

Why are we using the requirements applied to health labs? There’s no regulation for the use of mercury in research. In addition, the Brazilian law does not establish how should be the handling of toxic metals. It mentions only regarding to their levels released into the environment, and it is not very accurate. This brings us to a question, how to control the amount of mercury in the environment if there is no control in handling it. Due to lack of appropriate legislation in not only to dump mercury but also for use in research, the team UFAM_Brazil have been engaged in trying to improve this legislation. (read more).

Sanitary requirements for the handling of mercury in laboratories (Applied to health labs).

PPE

Masks , Gloves , Lab coat.

Equipments

Use masks and goggles to protect eyes, mouth and nose against volatile chemicals, splashes and inhalation of aerosol particles. Use an autoclave to make sterile infectious material by effective heat. Use of bottles with screw cap to prevent spills and aerosols. Countertops are decontaminated before and after the development of some activities. Before developing the experiment, UV light cabin remains on for 15 minutes. Alcohol and detergent are used as chemical agents for cleaning hands against microorganisms.

Floor and Bench

The floor of the room should be waterproof, without pores and cracks and easily cleanable. The work bench smooth and easy to clean.

Storage of the mercury

The storage of mercury should be in unbreakable and airtight jars. Code ADR: 8,66 To carry the mercury in a liquid state. Stored mercury must be labeled with the phrases: R23 (" Toxic by inhalation") and R33 ("Danger of cumulative effects"). Should also contain the phrases: S1/2 ("Keep under lock and keep out of reach of children"), S7 ("Keep container tightly sealed") and S45 ("In case of accident or sickness, immediately call your doctor (if possible show him the label).

Disposal of mercury

Disposal of waste mercury is considered biohazardous. Therefore, they must be placed in unbreakable bottles containing water inside and are sealed, labeled "biohazardous" and placed in a plastic bag milky white. All generated waste, such as cultures and stocks, microorganism culture media, the instruments used to inoculate cultures and biological samples are placed in order to be treated by a physical process to reduce or eliminate the microbial load. The sharps are discarded in rigid containers resistant to puncture, break and leak and properly autoclaved before disposal.

The MerColi

The American Biological Safety Association (ABSA) was founded in 1984 to promote biosafety as a scientific discipline and serve the growing needs of biosafety professionals throughout the world. The Association's goals are to provide a professional association that represents the interests and needs of practitioners of biological safety, and to provide a forum for the continued and timely exchange of biosafety information. Our project this year used E. coli of different strains: E. coli RR1, E. coli DH10B, E. coli DH5 alpha and E. coli JM110. All of those are included in risk group 1, according to ABSA, as it shown in the Table below:

Species name

Risk Group

How it was acquired

Disease risk to human

E coli RR1

1 (Link)

genes were synthesized

Highly unlikely to cause disease in healthy laboratory workers, animals. But may cause irritation to skin, eyes, and respiratory tract if the contact is direct

E coli DH10B

1 (Link)

Assembly

May cause irritation to skin, eyes, and respiratory tract

E coli DH5 alpha

1 (Link)

genes were synthesized

May cause irritation to skin, eyes, and respiratory tract

E coli JM110

1 (Link)

genes were synthesized + BBa_E0840 or BBa_K346004

Present no hazard to human health, non-pathogenic, non-toxicogenic. Cause irritation to skin, eyes, and respiratory tract if the contact is direct