What is the Effect of Microgravity on Bacterial Biofilm Formation on Soft Contact Lenses? Principal Investigator: Maya Karri, 5th Grade New Explorations into Science, Technology & Mathematics (NEST+m) New York City, NY Submitting Teacher Facilitators: Margaux Stevenson & Marvin Cadornigara, NEST+m Collaborators: (listed in alphabetical order): Alex Apelewicz, Chloe Coward, Tori Feinstein, Michelle Fridman, Jack Galligan, Anabel Giacobbi, Benjamin Goodstein, Quinci Huston, Andrew Kogan, Vivian Lee, Spring Lin, Haydn Long, Samantha Mayol, Ines Menendez, Fiona O’Reilly, Jean Carlos Paredes, Catherine Pyne, Kiara Reyes, Natalie Roston, Jacob Rubakha, Maximillian Shen, Rachel Svoyskiy, James Thompson, Abel Torres, Madison Williams, Danny Yang

Proposal Summary My proposal is about bacterial biofilm formation on soft contact lenses, which are made out of hydrogel. A bacterial biofilm is a colony of cells held together by a slimy substance that form on the surface of a contact lens. This is harmful because antibiotics, disinfectants, and host defenses do not work against them. In my experiment, I will be investigating how microgravity affects biofilm formation on soft contact lenses.

Experiment Materials and Handling Requirements Volume 1 (Contact Lens & Growth Medium) 2.8 mL of lactose broth CooperVision Contact Lens, Monthly Lenses, Prescription -2.5 Volume 2 (Bacteria & Growth Medium) 30 colonies of Serratia marcescens 2.8 mL ​Lactose broth ​ Volume 3 (Fixative) 2.8 mL 9% w/v paraformaldehyde ​Proposed Experiment Timeline of Crew Interactions Aboard ISS A=0 : Open Clamp A, Tilt Tube Once to Mix D-2 : Open Clamp B, Tilt Tube Once to Mix

Question to be Addressed The question being investigated is: What is the Effect of Microgravity on Bacterial Biofilm Formation on Soft Contact Lenses? Steps in Biofilm Formation: 1. Planktonic cells adhere to a surface 2. The cells form a microcolony 3. They release a slimy substance that holds them together 4. More bacterial cells join the colony 5. They form a mushroom-shaped film called a biofilm

Experimental Design Preparing the Setup ​In order for the bacteria to form, I must grow colonies for 24 hours at 37 degrees Celsius. 30 colonies must be collected for the ground experiment and 30 colonies for the experiment going to space. I will then fill the three sections of the FME tubes: first the contact lens with 2.8 mL of lactose broth, second the 30 colonies of Serratia marcescens in 2.8 mL of lactose broth, and third the fixative. The ground setup will be identical to that in microgravity. Analysis Once the FME returns, I will compare the biomass and the viability of the two lenses. I will measure the biomass using dry weight measurement. I will use a live/dead bacterial staining kit to measure the viability on each lens. After I stain the lenses, I will view them using a scanning electron microscope.

Letter of Certification by the Teacher Facilitator Acknowledgements I would like to thank my teacher, Ms. Stevenson; NEST+m’s SSEP program administrator and NEST+m Assistant Principal Mr. Brendan Alfieri; and NEST+m science teacher Mr. Marvin Cardornigara. I would also like to thank the SSEP workers, The Center for the Advancement of Science in Space, Subaru of America, Inc., The New York State Space Grant Consortium, the NYC Department of Education, and the NEST+m PTA. Letter of Certification by the Teacher Facilitator May 2015 I certify that the student team designed the experiment described herein and authored this proposal, and not a teacher, parent, or other adult. I recognize that the purpose of this letter is to ensure that there was no adult serving to lead experiment definition and design, or write the proposal, and thereby provide content and/or professional expertise beyond that expected of a student-designed and student-proposed experiment. I also understand that NCESSE recognizes that facilitation of thinking across the student team by the team’s Teacher Facilitator, and other teachers, parents, and local area researchers, is not only to be encouraged but is absolutely vital if students are to receive the necessary guidance on the process of scientific inquiry, experimental design, how to do background research in relevant science disciplines, and on writing the proposal. I also certify that the samples list and the special handling requests listed in this proposal are accurate and conform to the requirements for SSEP Mission 5 to ISS. I confirm that the team, after reviewing their procedure and budget for obtaining the samples for the experiment, is certain that they will be able to obtain the necessary samples for their experiment in time to meet the deadline for shipping the flight-ready FME to NanoRacks. If using human samples, the team is aware that these samples must be tested for prohibited viruses before the experiment can be selected for flight. Finally, the Teacher Facilitator certifies that the student team will have access to the proper facilities to prepare the Fluid Mixing Enclosure for flight and to analyze the samples after the flight. Margaux Stevenson Teacher Facilitator