Team:Debrecen Hungary/More

From 2011.igem.org

Revision as of 13:44, 21 September 2011

Safety

1. Would any of your project ideas raise safety issues in terms of researcher safety, public safety, or environmental safety?

No issue of researcher safety, public safety or environmental safety were raised during Debrecen's iGEM 2011 project. We only worked with non-hazardous, non-infectious, commonly used and accepted bacteria strain (DH5α) and mammalian cancer cell lines (COS-1). When working with toxic chemicals (e.g. ethidiumbromide or estrogen), nitrile gloves, and white coats were worn. All of the work was conducted in a biosafety level S1 laboratory. Rules of the best microbiological practices were applied.

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

All material handled or distributed are non-hazardous and non-infectious. It agrees with all safety standards requested biosafety level 1, therfore the project get a full supports for the work done by the iGEM team. We developed synthetic LBD's for use in mammalian cells in fusion parts. These parts are completely harmless, no matter what organism they are transformed / transfected into.

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

All of our work complies with the University of Debrecen's local biosafety and bioethics regulations. Our project leader and team instructors oversaw the biological safety issue throughout the whole working time this year. There is also a biosafety supervisor who supervises the overall laboratory work biosecurity-wise. All team members who worked at the bench this summer have had proper safety training. We received approval from all overseeing groups.

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?

An important tool to raise interest and knowledge in safety issues is education. Open source webpages containing regulations, new ideas, video tutorials etc. can be designed. Online forums, trainings, courses should be organized in order to share new issues between groups and experts.

Testing of arsenic biosensor

Overview

Arsenic is a serious groundwater contaminant of major public health significance. During the 1970s, due to a high incidence of waterborne disease, millions of tube wells were drilled in Bangladesh to provide drinking water. Unfortunately, years later, many instances of ill health were found to be associated with consumption of this water, and it became apparent that many of the wells had inadvertently been drilled through arsenic bearing sediments and that the water was contaminated with arsenic in the form of arsenate and arsenite anions. Arsenic is a serious poison, and chronic consumption leads to arsenicosis, with symptoms such as skin lesions and cancers. The WHO limit for arsenic in drinking water is 10 ppb, though a more relaxed limit of 50 ppb is still in operation in many countries. Many wells exceed these limits. Similar contaminated groundwater has also been found to occur in many other countries, and one estimate suggests that 100 million people worldwide may be at risk. There is therefore a need for methods for testing and monitoring arsenic levels. Various highly accurate laboratory methods are available, but for large scale use in less developed regions, it would be preferable to have simple and cheap test kits which can be used in the field.

In 2006, Team Edinburgh developed a genetically engineered machine which was able to detect arsenite/arsenate ions in water samples. To see the part visit the Registry of Standard Biological Parts here: [http://partsregistry.org/Part:BBa_J33203].

As the Edinburgh team tested the system only on distilled water samples spiked with sodium-arsenate, we contacted the team and offered our help to test the machine on real-world samples to prove that it works not only on artificial but on natural samples as well (with many different compounds that may interfere with the part's action).

As the South-East of Hungary is highly affected by the arsenic problem (there are wells in which the level of arsenic exceeds the allowed limit), we had the possibility to use real-world samples for testing. The current situation of the arseinc problem in the drinking water of the south east region of Hungary can be found here: [1]

Method of testing

The contact person of Team Edinburgh 2006 sent us the protocol should be used according to their experiments. We collected samples from two wells which were known to contain elevated arsenic concentration (Szegfű and Madách street, Békéscsaba, Hungary). Find these places using Google Maps: [http://maps.google.com/maps?f=q&source=s_q&hl=hu&q=5600+B%C3%A9k%C3%A9scsaba,+Szegf%C5%B1+utca,+Magyarorsz%C3%A1g&sll=37.09024,-96.503906&sspn=34.945679,92.285156&ie=UTF8&cd=1&geocode=FeAfyAIdRVlBAQ&split=0&hq=&hnear=5600+B%C3%A9k%C3%A9scsaba,+Szegf%C5%B1+utca,+Magyarorsz%C3%A1g&ll=46.666579,21.059632&spn=0.021204,0.044546&t=h&z=15], [http://maps.google.com/maps?f=q&source=s_q&hl=hu&geocode=&q=5600+B%C3%A9k%C3%A9scsaba,+Mad%C3%A1ch+utca,+Magyarorsz%C3%A1g&sll=46.666579,21.059632&sspn=0.021204,0.044546&g=5600+B%C3%A9k%C3%A9scsaba,+Szegf%C5%B1+utca,+Magyarorsz%C3%A1g&ie=UTF8&hq=&hnear=5600+B%C3%A9k%C3%A9scsaba,+Mad%C3%A1ch+utca,+Magyarorsz%C3%A1g&ll=46.666314,21.070104&spn=0.021204,0.044546&t=h&z=15]

We used DH5 alpha E. coli cell line with lacZDM15 mutation as host cells, than we prepared a medium consisting of peptone, yeast extract, NaCl, K2HPO4, NaHCO3, bromthymol blue, and autoclaved water samples of different dilutions. We spiked these media with the transformed cells, and after 24 hours of incubation at 37°C we received that the part works also on real world samples, not only on samples made of water and sodium-arsenate. We used reference dilutions made from sodium arsenate and MilliQ water to estimate the level of arsenic in our samples.

Results

The colour change of Bromothymol Blue seemed to be directly proportional to the concentration of arsenic. According to the reference dilutions made from Na-arsenate and MilliQ water the concentration of arsenic in our collected samples is at least 15x higher than the WHO recommended limit (Fig.1., Fig.2., Fig.3.).

Fig.1.: Arsenic reference dilutions for estimating arsenic concentration of real world samples after 24 hours of incubation.

Fig.2.: Dilutions of Szegfű street sample after 24 hours of incubation.

Fig.3.: Dilutions of Madách street sample after 24 hours of incubation.

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