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Biology Research Project on Effects of Nitrites on Zebrafish Earns Students a Society of Toxicology Award

Posing in the laboratory were, from left, Ida Karimi, Alison Simmons, and Tom Simmons, advisor on the students' research project. (Keith Boyer photo)Two Indiana Area Senior High School students are getting a head start on their academic careers.

Ida Karimi and Alison Simmons, students at IUP during the summer, won the 2011 Mary Anne Stock Student Research Award from the Allegheny-Erie chapter of the Society of Toxicology for their research project “Effects of Nitrite on Development of Embryos and Early Larval Stages of the Zebrafish (Danio rerio).”

The proposal, reviewed by toxicologists, was praised as “a very cogent and well-written proposal with high and direct relevance to environmental toxicology.”

Karimi, the daughter of Roya and Majid Karimi, and Simmons, the daughter of Anne and Tom Simmons, will return to Indiana Area Senior High School in the fall to complete their senior years.

“We were lab partners at Indiana High School in Mr. [Bill] Warwick’s class for Advanced Placement biology in eleventh grade, and we decided we wanted to do something more this summer with a biology research project,” Simmons said. Both plan to major in biology after high school.

To complete the project, they studied under Simmons’ father, Tom, a faculty member in the Biology Department. Karimi’s father, Majid, is a faculty member in the Physics Department.

The project, which met all requirements of an IUP research project, including guidelines of the Institutional Animal Care and Use Committee, was designed to see if nitrites in high concentrations, which are known to cause human infantile methemoglobinemia (also known as blue baby syndrome, a condition resulting in oxygen deprivation in newborns) would cause malformations in developing zebrafish. The students chose zebrafish for the study because they are the standard vertebrate models for studying those issues.

Nitrate, which is converted into nitrite, is a compound found throughout the environment, and humans are exposed via ingestion of water and food. The students found that there are no published studies on the direct developmental effects of nitrites.

“We were really surprised to win the award, especially because we were competing against undergraduate and graduate students,” Karimi said.

The students were responsible for all aspects of the research project, including establishing a breeding zebrafish colony in an IUP laboratory.

“We didn’t count on the project being so time consuming,” Alison Simmons said. “There was a lot of cleaning when we first got started, and it was hard to get the environment exactly right for the fish.”

“At first, the fish kept dying, but eventually, we actually got more eggs than we expected, almost three hundred to four hundred eggs a day from each tank.”

The students then gradually exposed the fish eggs to increased concentrations of nitrites to determine when the larvae were affected by the toxin.

After six weeks of twenty hours a week of research, documentation, and study, the students found that nitrite exposure to the fish larvae was just as damaging in terms of developmental abnormalities as ethanol (alcohol), which, at high levels, has been documented as causing birth defects in humans.

“The students did a great job on the project,” Tom Simmons said. “It’s definitely a study that other researchers can use as a basis for ongoing research. The next step is determining if the concentration of nitrites that caused the developmental abnormalities in the zebrafish is a realistic model for humans. More study also needs to be done to analyze longer exposures to nitrites at lower concentrations and its effects on the zebrafish larvae.”

As part of their award-winning proposal, the students will present their findings at the spring meeting of the Allegheny-Erie chapter of the Society of Toxicology.