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Orlando Melendez

Publications and source records attributed to Orlando Melendez.

8 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS2017

A PCR Based Microbial Monitoring Alternative Method of Detection and Identification of Microbes Aboard ISS

Previous research has shown that microorganisms and potential human pathogens have been detected on the International Space Station (ISS) with additional introduction of new microflora occurring with every exchange of crew or addition of equipment and supplies. These microbes are readily transferred between crew and subsystems (i.e. ECLSS, environmental control and life support systems). As this can be detrimental to astronaut health and optimal performance of ISS systems, monitoring of systems such as ECLSS to include identification of microbial contaminants could prevent adverse effects on human health and life support systems. Current monitoring on ISS is laborious and utilizes culture based methods followed by sample return to Earth for complete analysis. Future, long-distance spaceflight missions will require real-time monitoring capabilities that enable efficient and rapid assessments of the microbial environment allowing for expedited decisions and more targeted response to cope with anomalies. Polymerase chain reaction (PCR), a molecular microbial monitoring method was chosen and numerous PCR instruments investigated for their potential to perform in microgravity conditions. Using ISS as a test bed for PCR verification in microgravity will enable NASA to assess whether molecular based microbiological sensors may be components of reliable, closed-loop life support and habitation systems in spacecraft, enhancing infrastructure capabilities through increased efficiency, reliability, and time savings by enabling sample analysis on orbit. NASA selected the Water Monitoring Suite as one of the rapid spaceflight hardware demonstration activities utilizing a streamlined process to minimize the time required to fly experimental flight hardware. The RAZOR EX (BioFire Defense, Salt Lake City, UT) system was part of the water monitoring suite and is a commercial off-the-shelf (COTS) real-time PCR instrument designed for field work. The RAZOR EX was originally designed for Department of Defense (DoD) under a small business innovative research (SBIR) grant and is ruggedized, compact and provides a rapid, sample to answer in less than an hour. PCR assays using a fluorescent probe were optimized and spiked with known concentrations of DNA (Pseudomonas aeruginosa) ranging from 0.002 to 20 ng. PCR reagents were lyophilized and configured in customized pouches and tested for flight readiness. Three types of water were used to rehydrate the reagents and demonstrate the fidelity of the PCR reaction in microgravity. Molecular grade deionized water served as a control while filtered and unfiltered ISS potable water served to test for chemical or biological inhibitors. All three types were compared to parallel ground test results. Nine tests were run on ISS (3 of each water type) and the critical threshold cycle (Ct) was compared to parallel ground tests completed at Kennedy Space Center, FL and Johnson Space Center, TX. All concentrations of Pseudomonas aeruginosa DNA were detected. A comparison of the Ct produced in real time PCR indicated similarity between flight and ground samples. There appeared to be no significant difference between flight or ground PCR reactions or between any of the three water types. This testing demonstrated the ability to perform molecular testing during spaceflight operations with similar sensitivity. It will allow for future ground development of molecular protocols and minimize the need for spaceflight testing. Future testing will include development of additional targets including environmental and health related organisms.

Christina Khodadad↗
NASA NTRS1997

Evaluation of AK-225(R), Vertrel(R) MCA and HFE A 7100 as Alternative Solvents for Precision Cleaning and Verification Technology

The National Aeronautics and Space Administration (NASA), Kennedy Space Center (KSC) Materials Science Division conducted a study to evaluate alternative solvents for CFC-113 in precision cleaning and verification on typical samples that are used in the KSC environment. The effects of AK-225(R), Vertrel(R), MCA, and HFE A 7100 on selected metal and polymer materials were studied over 1, 7 and 30 day test times. This report addresses a study on the compatibility aspects of replacement solvents for materials in aerospace applications.

Orlando Melendez↗
NASA NTRSDate not supplied

ISS Wastewater Pretreatment via DNA Pattern Picofilter Using Inorganic Brine Stimulant

Astronauts in the International Space Station (ISS) need as much water as they can access, whether it comes from people's breath, sweat or urine, recycled shower water or hand washing. It is of vital importance that any type of water is recycled and filtrated through different methods so it can be re-used. Recently, Cerahelix Inc. offered a ceramic tubular membrane element that uses DNA strands as a pattern in a sol-gel process that allows the sintered product to have a pore size in the picometer scale and claims to achieve ten times higher purity than other commercially available ceramic filters. This should allow higher purities and yields at reduced energy costs and theoretically allow near total dewatering of the reject stream and greater than 80% removal of polyvalent ions. Trials were performed with Cerahelix Inc. filters using an inorganic brine simulant at two pHs to test the efficiency of Pico Helix TM membranes and determine their feasibility for spacecraft wastewater treatment processes. Emphasis was placed on the polyvalent ions: SO42-, PO43-, Mg2+, and Ca2+. These ions should be rejected almost entirely from the feed solution and, not permeate through the filter. Results show that at a pH of 4.2, permeation of the polyvalent ions varied from 59% to 74% and at pH of 8.0, permeation varied from 72% to 87%. An Extended Nernst Planck (ENP) approach that describes the mass transfer process for a pico-filtration membrane was used to construct a model to inform the experimental expectations and outcome.

Carolina Franco↗