Environmental Sciences Fair Project
Toxicity of Carbon Nanotubes in Daphnia magna

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Project Information
Title: Toxicity of Carbon Nanotubes in Daphnia magna
Subject: Environmental Sciences
Grade level: High School - Grades 10-12
Academic Level: Advanced
Project Type: Experimental
Cost: Medium
2nd place, Canada Wide Virtual Science Fair ($100)
WWSEF Life Science Gold Medal
Affiliation: Canada Wide Virtual Science Fair
Year: 2010
Description: Daphnia magna neonates were exposed to solutions of carbon nanotubes and humic acid to determine the acute toxicity of carbon nanotubes in combination with humic acid. Spectrophotometry analysis was also employed to evaluate the effects of humic acid on carbon nanotube solubility.
Link: www.virtualsciencefair.org...

Carbon Nanotubes

Carbon nanotubes (CNTs; also known as buckytubes) are allotropes of carbon with a cylindrical nanostructure. Nanotubes have been constructed with length-to-diameter ratio of up to 132,000,000:1, significantly larger than any other material. These cylindrical carbon molecules have novel properties, making them potentially useful in many applications in nanotechnology, electronics, optics, and other fields of materials science, as well as potential uses in architectural fields. They may also have applications in the construction of body armor. They exhibit extraordinary strength and unique electrical properties, and are efficient thermal conductors.

Nanotubes are members of the fullerene structural family, which also includes the spherical buckyballs. The ends of a nanotube may be capped with a hemisphere of the buckyball structure. Their name is derived from their size, since the diameter of a nanotube is on the order of a few nanometers (approximately 1/50,000th of the width of a human hair), while they can be up to 18 centimeters in length (as of 2010). Nanotubes are categorized as single-walled nanotubes (SWNTs) and multi-walled nanotubes (MWNTs).

Applied quantum chemistry, specifically, orbital hybridization best describes chemical bonding in nanotubes. The chemical bonding of nanotubes is composed entirely of sp2 bonds, similar to those of graphite. These bonds, which are stronger than the sp3 bonds found in diamonds, provide nanotubules with their unique strength. Moreover, nanotubes naturally align themselves into "ropes" held together by van der Waals forces.

Toxicity of Carbon Nanotubes

Determining the toxicity of carbon nanotubes has been one of the most pressing questions in nanotechnology. Unfortunately, such research has only just begun. Thus, the data are still fragmentary and subject to criticism. Preliminary results highlight the difficulties in evaluating the toxicity of this heterogeneous material. Parameters such as structure, size distribution, surface area, surface chemistry, surface charge, and agglomeration state as well as purity of the samples, have considerable impact on the reactivity of carbon nanotubes. However, available data clearly show that, under some conditions, nanotubes can cross membrane barriers, which suggests that if raw materials reach the organs they can induce harmful effects such as inflammatory and fibrotic reactions.

A study led by Alexandra Porter from the University of Cambridge shows that CNTs can enter human cells and accumulate in the cytoplasm, causing cell death.

Results of rodent studies collectively show that regardless of the process by which CNTs were synthesized and the types and amounts of metals they contained, CNTs were capable of producing inflammation, epithelioid granulomas (microscopic nodules), fibrosis, and biochemical/toxicological changes in the lungs. Comparative toxicity studies in which mice were given equal weights of test materials showed that SWCNTs were more toxic than quartz, which is considered a serious occupational health hazard when chronically inhaled. As a control, ultrafine carbon black was shown to produce minimal lung responses.

The needle-like fiber shape of CNTs, similar to asbestos fibers, raises fears that widespread use of carbon nanotubes may lead to mesothelioma, cancer of the lining of the lungs often caused by exposure to asbestos. A recently-published pilot study supports this prediction. Scientists exposed the mesothelial lining of the body cavity of mice, as a surrogate for the mesothelial lining of the chest cavity, to long multiwalled carbon nanotubes and observed asbestos-like, length-dependent, pathogenic behavior which included inflammation and formation of lesions known as granulomas. Authors of the study conclude:

"This is of considerable importance, because research and business communities continue to invest heavily in carbon nanotubes for a wide range of products under the assumption that they are no more hazardous than graphite. Our results suggest the need for further research and great caution before introducing such products into the market if long-term harm is to be avoided."

According to co-author Dr. Andrew Maynard:

"This study is exactly the kind of strategic, highly focused research needed to ensure the safe and responsible development of nanotechnology. It looks at a specific nanoscale material expected to have widespread commercial applications and asks specific questions about a specific health hazard. Even though scientists have been raising concerns about the safety of long, thin carbon nanotubes for over a decade, none of the research needs in the current U.S. federal nanotechnology environment, health and safety risk research strategy address this question."

Although further research is required, results presented today clearly demonstrate that, under certain conditions, especially those involving chronic exposure, carbon nanotubes can pose a serious risk to human health.

Daphnia Magna and Ecotoxicity Testing

Daphnia magna is a species of Daphnia (a cladoceran freshwater water flea) which is native to northern and western North America. It is also widely distributed in Eurasia and in some regions of Africa.

It is widely used as a laboratory animal for testing ecotoxicity starting with Einar Naumann in 1934.

The use of Daphnia magna as an experimental animal for such purposes is advantageous in many respects. Daphnids are small, reaching a size of five mm, so that a great many can be reared in a small space. They have a relatively short life span, which reaches a maximum of about two months when they are reared at 25C. Daphnids are easy to culture, requiring only water containing bacteria or their equivalent for food. They can be grown individually in small bottles or in mass culture in large aquaria. They mature early, giving birth to young within their first week of life. After their first brood, they give rise to new broods every two or three days throughout the remainder of their lives. An average of twenty or more young may be produced in each brood. Each female who lives to a ripe old age can bear four hundred or more offspring. Again, all the young from one female are genetically like the mother if produced parthenogenically, and reproduction can be limited to parthenogenesis if the proper conditions are maintained. Further, daphnids are representatives of a class of animals that serve as food for many fish, especially while the fish are young. Fishes do not remain long in waters where their food supply has been destroyed, even though the fishes may not be affected directly. For these reasons daphnids should prove satisfactory for testing waters for toxic materials.

D. magna is specified to be used in the OECD Guidelines for the Testing of Chemicals, Tests No. 202 "Daphnia sp., Acute Immobilisation Test and Reproduction Test", and Test No. 211 "Daphnia magna Reproduction Test". Test No. 202 is a 48 hour acute toxicity study, where young Daphnia are exposed to varying concentrations of the substance under test and the EC50 determined. Other Daphnia species than D. magna may occasionally be used, but labs mostly use D. magna as standard.

Test No. 211 is a 21 day chronic toxicity test, at the end of which, the total number of living offspring produced per parent animal alive at the end of the test is assessed, in order to determine the lowest observed effect concentration of the test substance.

For More Information:
Carbon nanotube
Daphnia Magna

Source: Wikipedia (All text is available under the terms of the Creative Commons Attribution-ShareAlike License)

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