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Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2023 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2024  
Task Last Updated: 02/13/2023 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Griko, Yuri  
Center Contact: 650-604-0519 
Yuri.V.Griko@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program: ISS 
No. of Post Docs:  
No. of PhD Candidates:
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Flight Assignment/Project Notes: NOTE: End date changed to 04/05/2024 per NSSC information (Ed., 2/22/23)

Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and European Space Agency (ESA)’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits: Microbial biofilms are known to cause persistent infections as well as degrade a variety of materials including metals. Biofilms are notorious for their persistence and resistance to eradication. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The objectives of this project include determining the efficacy of biofilm inhibition by different oxidation states of metals and inhibition by nanoscale texture patterns on various metals. The results from the nano-scale texture patterns represent a new technology that is applicable to inhibiting biofilm formation in hospitals, and also in the pharmaceutical and industries where biofilm corrosion is a problem.

Task Progress & Bibliography Information FY2023 
Task Progress: For the BIOFILMS experiment, copper and brass (CuZn37) were selected as antimicrobial surfaces, while stainless steel is used as the inert reference. All three surfaces are tested with and without tailored surface functionalization (that is, patterned grooves laser cut into the metal surfaces at 3 micron and 800 nm levels). Using the KUBIK facility in the Columbus laboratory on board the ISS, the antimicrobial efficacy of the surfaces is being tested under three different gravity levels (µg, 0.4 x g, 1 x g). [Ed. Note: KUBIK is a small incubator developed by the European Space Agency (ESA) for self-contained microgravity experiments on board the International Space Station (ISS).] Staphylococcus capitis, Cupriavidus metallidurans, and Acinetobacter radioresistens were selected as bacterial model organisms. The bacteria are incubated in specialized hardware that allows biofilm formation to occur under controlled conditions while being exposed to the different metal surfaces. The BIOFILMS experiment is being conducted during three flights on board the ISS to accommodate the complete set of sample configurations (metal type, surface functionalization, bacteria, and gravity level). The first launch of BIOFILMS was in August 2021 with SpX 23, the second in July 2022 with SpX 25, and the last flight is planned for March 2023 with SpX 27. The following are preliminary results from the first flight and part of the second, with the caveat that a complete interpretation and evaluation of the data can only be made after the completion of all three flights and post flight evaluations. The results of BIOFILMS will help to make space travel safer and more sustainable by providing new insights and design capabilities in contamination control.

The Biofilms 1 data analysis is complete. It was found that patterning at both the 3 micron and 800 nm levels seems to stress the organisms, as evidenced by the production of more extracellular material (presumably mostly extracellular polysaccharides) on all metal surfaces tested. Copper shows clear inhibition of microbial growth on all types of surfaces. The brass coupon appears to inhibit microbial growth but somewhat less than copper (quantification requires further analyses). The effect of gravity is not readily apparent and appears to be masked by the effects of the metals and functionalization. It is clear that more detailed analyses need to be done. The extracellular material produced on the copper and brass plates has a more spotty/blotchy appearance than the extracellular polymeric substances (EPS) on the stainless steel plates. This might be due to the interaction of copper with the EPS. It was decided that Energy Dispersive X-Ray Analysis (EDAX) would be a good way to discern the differences and similarities. Biofilms 2 launched on SpX-25 at the end of July and we are currently working on those samples. Preliminary results appear to be similar to those from Biofilms 1. Biofilms 3 is scheduled to launch in February/March 2023.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
Abstracts for Journals and Proceedings Siems K, Müller D, Ahmed A, Van Houdt R, Mancinelli RL, Brix K, Kautenburger R, Krause J, Vukich M, Tortor A, Roesch C, Holland G, Laue M, Mücklich F, Moeller R. "Update on the spaceflight experiment “BIOFILMS”: Testing laser-structured antimicrobial surfaces under space conditions." 38th Annual Meeting of the American Society for Gravitational and Space Research, Houston, TX, November 9-12, 2022.

Abstracts. 38th Annual Meeting of the American Society for Gravitational and Space Research, Houston, TX, November 9-12, 2022. , Nov-2022

Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2022 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2023  
Task Last Updated: 03/02/2022 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Griko, Yuri  
Center Contact: 650-604-0519 
Yuri.V.Griko@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program: ISS 
No. of Post Docs:  
No. of PhD Candidates:
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and European Space Agency (ESA)’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits: Microbial biofilms are known to cause persistent infections as well as degrade a variety of materials including metals. Biofilms are notorious for their persistence and resistance to eradication. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The objectives of this project include determining the efficacy of biofilm inhibition by different oxidation states of metals and inhibition by nanoscale texture patterns on various metals. The results from the nano-scale texture patterns represent a new technology that is applicable to inhibiting biofilm formation in hospitals, and also in the pharmaceutical and industries where biofilm corrosion is a problem.

Task Progress & Bibliography Information FY2022 
Task Progress: Progress in 2021

BIOFILMS #1. The assembly of the BIOFILMS hardware (HW), including all the preparation, shipping and logistics, for the first flight was completed and went well. The first flight occurred on Aug 29, 2021 (0714 GMT) with Space-X CRS 23 (SpX23) with our BIOFILMS HW on board. The hardware was successfully loaded into KUBIK and the experiment was conducted successfully. [Ed. Note: KUBIK is a small incubator developed by the European Space Agency (ESA) for self-contained microgravity experiments on board the International Space Station (ISS).] The samples were fixed and returned to Earth. On October 17, 2021, the samples arrived at the German Aerospace Center (DLR) and the analysis of the flight samples began. The analysis and data interpretation are ongoing. The results of the BIOFILMS 1 flight will be provided after the analyses and interpretation of the data have been completed. Additionally, a postflight debriefing meeting with ESA was held.

BIOFILMS #2. In addition to analyzing the samples from BIOFILMS 1, we are preparing for the BIOFILMS 2 flight in coordination with ESA and Keyser Italia. BIOFILMS 2 is now targeting SpX-25, planned for launch during the 4th week of May 2022. This decision was driven by the potential risk linked with the time window available for BIOFILMS 2 readiness, squeezed to 2 months by SpX-23 and SpX-24 launch dates. Moreover, ESA was informed by Space-X that cold stowage capabilities on SpX-24 were overly subscribed and no room would have been available for BIOFILMS 2. We had a similar risk for SpX-23 that would have required a re-design of the BIOFILMS packaging in order to fit the HW in only one Double Cold Bag (DCB). SpX-25 and SpX-26 launch dates are only three months apart; that is, SpX-25 is scheduled to return during the 2nd week of July and SpX-26 is scheduled to launch during the 4th week of October. ESA has stated that it is too early to get to a definitive date for BIOFILMS 3.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
Articles in Peer-reviewed Journals Siems K, Müller DW, Maertens L, Ahmed A, Van Houdt R, Mancinelli RL, Baur S, Brix K, Kautenburger R, Caplin N, Krause J, Demets R, Vukich M, Tortora A, Roesch C, Holland G, Laue M, Mücklich F, Moeller R. "Testing laser-structured antimicrobial surfaces under space conditions: The design of the ISS experiment BIOFILMS." Front. Space Technol. 2022 Jan 3;2. https://doi.org/10.3389/frspt.2021.773244 , Jan-2022
Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2021 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2023  
Task Last Updated: 03/09/2021 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Griko, Yuri  
Center Contact: 650-604-0519 
Yuri.V.Griko@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program: ISS 
No. of Post Docs:  
No. of PhD Candidates:
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and European Space Agency (ESA)’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits: Microbial biofilms are known to cause persistent infections as well as degrade a variety of materials including metals. Biofilms are notorious for their persistence and resistance to eradication. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The objectives of this project include determining the efficacy of biofilm inhibition by different oxidation states of metals and inhibition by nanoscale texture patterns on various metals. The results from the nano-scale texture patterns represent a new technology that is applicable to inhibiting biofilm formation in hospitals, and also in the pharmaceutical and industries where biofilm corrosion is a problem.

Task Progress & Bibliography Information FY2021 
Task Progress: Progress in 2020

Testing antimicrobial surfaces in BIOFILMS hardware. Utilizing the BIOFILMS hardware allows for the analysis of bacterial biofilms and the influence of antimicrobial surfaces in different gravitational regimes. Here, two models of the BIOFILMS hardware were tested. First, a reduced model (RM) was tested for proof of functionality. Second, a scientific model (SM) was tested. The SM is similar in structure and function to the flight model, that will be used in aboard the ISS. Common features of all BIOFILMS hardware models are that they contain sample plates, manufactured with different antimicrobial surfaces, and a microbial growth chamber. The growth chamber enables direct contact between the microbe and the antimicrobial surface, and permits gas exchange through a gas permeable membrane.

Three different types of metals were evaluated as sample plates in the BIOFILMS hardware. They were tested for their ability to prevent microbial growth and thus biofilm formation. Steel was used as reference material because it is not known to have antimicrobial properties. Pure copper and brass as an alloy of copper, containing 63% copper and 37% zinc, were tested as antimicrobial surfaces. The sample plates had a size of 10 mm x 25 mm. In addition to testing smooth surfaces, structured surfaces generated by direct laser interference patterning (DLIP) were also tested. The created structures were 3µm in size.

Results: Testing BIOFILMS spaceflight hardware. The tests served to ensure the functionality of the hardware and to evaluate the influence of different antimicrobial surfaces on growth and biofilm formation of S. capitis K1-2-2-23. First, a biocompatibility test was performed to determine if all of the built-in components were suitable for conducting microbiological experiments. Following that, the reduced model (RM) and the scientific model (SM) of the BIOFILMS hardware were tested. The tests were all successful. The bio compatibility testing showed that the materials used were not inhibitory. The results showed that within the configuration for the flight experiment the copper and brass materials were inhibitory compared to the stainless steel controls.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
Articles in Peer-reviewed Journals Alder-Rangel A, Idnurm A, Brand AC, Brown AJP, Gorbushina A, Kelliher CM, Campos CB, Levin DE, Bell-Pedersen D, Dadachova E, Bauer FF, Gadd GM, Braus GH, Braga GUL, Brancini GTP, Walker GM, Druzhinina I, Pócsi I, Dijksterhuis J, Aguirre J, Hallsworth JE, Schumacher J, Wong KH, Selbmann L, Corrochano LM, Kupiec M, Momany M, Molin M, Requena N, Yarden O, Cordero RJB, Fischer R, Pascon RC, Mancinelli RL, Emri T, Basso TO, Rangel DEN. "The Third International Symposium on Fungal Stress - ISFUS." Fungal Biol. 2020 May;124(5):235-52. https://doi.org/10.1016/j.funbio.2020.02.007 ; PMID: 32389286; PMCID: PMC7438019 , May-2020
Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2020 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2023  
Task Last Updated: 02/07/2020 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Griko, Yuri  
Center Contact: 650-604-0519 
Yuri.V.Griko@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program: ISS 
No. of Post Docs:  
No. of PhD Candidates:
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and European Space Agency (ESA)’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits: Microbial biofilms are known to cause persistent infections as well as degrade a variety of materials including metals. Biofilms are notorious for their persistence and resistance to eradication. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The objectives of this project include determining the efficacy of biofilm inhibition by different oxidation states of metals and inhibition by nanoscale texture patterns on various metals. The results from the nano-scale texture patterns represent a new technology that is applicable to inhibiting biofilm formation in hospitals, and also in the pharmaceutical and industries where biofilm corrosion is a problem.

Task Progress & Bibliography Information FY2020 
Task Progress: Because of its fast generation time we used Vibrio natriegens as a model test organism for a variety of space environment related studies where generation time is a critical factor. Specifically, V. natriegens was used as a tool to study growth characteristics by determining the viable cell number and antibiotic susceptibility under simulated microgravity using a 2D clinostat (60 rpm) to establish a test system that resolves changes in microbial growth on a solid surface (agar) under microgravity. The data show that V. natriegens biomass increases significantly after 24 h at 37°C under simulated microgravity. The final cell population after cultivation under simulated microgravity was 60-fold greater than when cultivated under normal terrestrial gravity (1 x g). No change in susceptibility to the antibiotic rifampicin after cultivation under simulated microgravity or normal gravity was detected. These data show that V. natriegens is a new and innovative model organism for microbial microgravity research.

Preparative work for the upcoming EST & science verification test (SVT) tests was conducted. These tests included performing laboratory microbiology & analytical testing for sample preparation, cultivation, fixation, and potential post-flight analyses.

Desiccation testing was one important part of the microbiological testing. To complete this test, samples of the ISS isolate Staphylococcus capitis subsp. capitis K1-2-2-23 and the wild type (DSM 20326) were grown to mid-log phase and washed by centrifugation. The pellets were resuspended in buffered saline to a pre-determined cell concentration, removed from the centrifugation, and aliquots dried in Eppendorf tubes. Periodically, samples of the dried organisms were tested for viability with respect to desiccation tolerance). These data from these tests illustrated that the ISS isolate is more tolerant to desiccation than the wild type.

Additional tests were conducted to determine the viability of Staphylococcus capitis subsp. capitis K1-2-2-23 compared to the wild type (DSM 20326) when exposed to increasing levels of hydrogen peroxide. Hydrogen peroxide is an oxidizing agent that creates radical oxygen species in the cell that can kill cells and interfere with biofilm formation. The results showed that the ISS isolate is more tolerant to this oxidizing agent.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
Abstracts for Journals and Proceedings Siems K, Mancinelli RL, Moeller R, et al. "Staphylococcus capitis ISS isolate as a model organism for evaluating antimicrobial surfaces within the upcoming space flight experiment BIOFILMS." 35th Annual Meeting of the American Society for Gravitational and Space Research, Denver, CO, November 20-23, 2019.

Program and Abstracts. 35th Annual Meeting of the American Society for Gravitational and Space Research, Denver, CO, November 20-23, 2019. , Nov-2019

Abstracts for Journals and Proceedings Mancinelli RL, Cortesao M, Moeller R. "Microbes in Space: An overview." International Symposium on Fungi/Microbes Under Stress, San Jose de Los Campos, Brazil, May 18-26, 2019.

Symposium Abstract Book, International Symposium on Fungi/Microbes Under Stress, San Jose de Los Campos, Brazil, May 18-26, 2019. , May-2019

Articles in Peer-reviewed Journals Garschagen LS, Mancinelli RL, Moeller R. "Introducing Vibrio natriegens as a microbial model organism for microgravity research." Astrobiology. 2019 Oct;19(10):1211-20. https://doi.org/10.1089/ast.2018.2010 ; PubMed PMID: 31486680 , Oct-2019
Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2019 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2023  
Task Last Updated: 02/08/2019 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Sato, Kevin  
Center Contact: 650-604-1104 
kevin.y.sato@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program:  
No. of Post Docs:  
No. of PhD Candidates:  
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and ESA’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits: Microbial biofilms are known to cause persistent infections as well as degrade a variety of materials including metals. Biofilms are notorious for their persistence and resistance to eradication. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The objectives of this project include determining the efficacy of biofilm inhibition by different oxidation states of metals and inhibition by nanoscale texture patterns on various metals. The results from the nano-scale texture patterns represent a new technology that is applicable to inhibiting biofilm formation in hospitals, and also in the pharmaceutical and industries where biofilm corrosion is a problem.

Task Progress & Bibliography Information FY2019 
Task Progress: The efficacy of wetted oxidized copper layers and pure copper surfaces as antimicrobial agents was tested by applying cultures of Escherichia coli and Staphylococcus cohnii to these metallic surfaces. Stainless steel surfaces were used as non-inhibitory control surfaces. The production of reactive oxygen species and membrane damage increased rapidly within 1 h of exposure on pure copper surfaces, but the effect on cell survival was negligible even after 2 h of exposure. However, longer exposure times of up to 4 h led to a rapid decrease in cell survival, whereby the survival of cells was additionally dependent on the exposed cell density. Finally, the release of metal ions was determined to identify a possible correlation between copper ions in suspension and cell survival. These measurements indicated a steady increase of free copper ions, which were released indirectly by cells presumably through excreted complexing agents. These data indicate that the application of antimicrobial surfaces in spaceflight facilities could improve crew health and mitigate material damage caused by microbial contamination and biofilm formation. Furthermore, the results of this study indicate that cuprous oxide layers are superior to pure copper surfaces related to the antimicrobial effect and that cell density is a significant factor that influences the time dependence of antimicrobial activity.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
Articles in Peer-reviewed Journals Hahn C, Hans M, Hein C, Mancinelli RL, Mucklich F, Wirth R, Rettberg P, Hellweg CE, Moeller R. "Pure and oxidized copper materials as potential antimicrobial surfaces for spaceflight activities." Astrobiology. 2017 Dec;17(12):1183-91. https://doi.org/10.1089/ast.2016.1620 ; PubMed PMID: 29116818 , Dec-2017
Project Title:  BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation Reduce
Images: icon  Fiscal Year: FY 2018 
Division: Space Biology 
Research Discipline/Element:
Space Biology: Cell & Molecular Biology   | Microbiology  
Start Date: 04/06/2018  
End Date: 04/05/2023  
Task Last Updated: 07/11/2018 
Download report in PDF pdf
Principal Investigator/Affiliation:   Mancinelli, Rocco  Ph.D. / Bay Area Environmental Research (BAER) Institute 
Address:  Mail Stop 239-4, NASA Ames Research Center 
 
Moffett Field , CA 94035 
Email: mancinelli@baeri.org 
Phone: (650) 604-6165  
Congressional District: 18 
Web:  
Organization Type: NON-PROFIT 
Organization Name: Bay Area Environmental Research (BAER) Institute 
Joint Agency:  
Comments:  
Co-Investigator(s)
Affiliation: 
Möller, Ralf  Ph.D. Principal Investigator--German Aerospace Center (DLR e.V.) 
Key Personnel Changes / Previous PI: Rocco L. Mancinelli, Ph.D., is U.S. Co-Investigator on this German Aerospace Center (DLR), Institute of Aerospace Medicine project. Principal Investigator is Ralf Möller, Ph.D., German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology Department.
Project Information: Grant/Contract No. 80NSSC18K0751 
Responsible Center: NASA ARC 
Grant Monitor: Sato, Kevin  
Center Contact: 650-604-1104 
kevin.y.sato@nasa.gov 
Unique ID: 11779 
Solicitation / Funding Source: 2014 ILSRA--Flight Opportunities for Space Life Sciences (non-US proposers) 
Grant/Contract No.: 80NSSC18K0751 
Project Type: FLIGHT,GROUND 
Flight Program:  
No. of Post Docs:  
No. of PhD Candidates:  
No. of Master's Candidates:  
No. of Bachelor's Candidates:  
No. of PhD Degrees:  
No. of Master's Degrees:  
No. of Bachelor's Degrees:  
Space Biology Element: (1) Cell & Molecular Biology
(2) Microbiology
Space Biology Cross-Element Discipline: (1) Reproductive Biology
Space Biology Special Category: (1) Cell Culture
(2) Translational (Countermeasure) Potential
(3) Bioregenerative Life Support
Task Description: Funding is for Dr. Rocco Mancinelli's role as U.S. Co-Investigator for this German Aerospace Center (DLR), Institute of Aerospace Medicine project, "BIOFILMS: Testing the Efficacy of Biofilm Formation by Antimicrobial Metal Surfaces under Spaceflight Conditions - An Effective Strategy to Prevent Microbial Biofilm Formation."

As Co-Investigator on the project, Dr. Mancinelli will provide his experience and expertise in microbiology and spaceflight to help design the flight experiment as well as the ground controls. He will also help trouble-shoot the system should it be necessary. He will play a major role in data interpretation, data analysis, and data management. He will help guide the ground control design and construction both on site (at the DLR) as well as remotely at NASA Ames. In addition, Mancinelli will take the lead in developing a conceptual model describing the effects of micro-gravity on the growth and development of biofilms as well as for the biofilms grown on metallic inhibitor surfaces.

To achieve many of the goals of NASA’s and ESA’s space programs requires an enduring human presence in space. Long term human missions require sustained crew health and safety. A research area that is important in sustaining crew health is the development of improved spaceflight-suitable methods for microbiological monitoring, as well as contamination control and reduction. The International Space Station (ISS) is a confined and isolated habitat in an extreme, hostile environment. The human and habitat microflora varies in response to changes in environmental conditions aboard the ISS. Changes in the microflora may result in an increased health risk for the crew. Microorganisms including microbial biofilms have been found on various habitat surfaces, inside the air and water handling systems as well as the hardware used on the ISS. Biofilms are known to cause damage to equipment from polymer deterioration, metal corrosion, and bio-fouling. The primary concern regarding crew health is characterized by activity of opportunistic pathogenic microorganisms that have been noted to accumulate in the closed environments of the ISS and other spacecraft on long-duration missions. Understanding the effects of the space environment, especially altered gravity, on microbial biofilms is crucial for the success of long-term human space missions. Surface-associated biofilm communities were abundant on the Mir space station and continue to be a challenge on the ISS. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of human immune function observed during spaceflight. Various studies have shown that certain metals reduce the number of contact-mediated microbial infections. Antimicrobial surfaces are defined as materials that contain an antimicrobial agent (such as silver, copper, and their alloys) that inhibits or reduces the ability of microorganisms to grow on the surface of a material. Antimicrobial surfaces are functionalized in a variety of different processes. The introduction of antimicrobial surfaces for medical, pharmaceutical, and industrial purposes has shown their unique potential for reducing and preventing microbial contamination. The contact killing of several types of microorganisms by copper has been assessed in multiple laboratory in-vitro studies. For sustained crew health and safety additional studies on the mechanisms involved in the formation of microbial biofilms and their efficient destruction under spaceflight conditions, i.e., long-term growth and adaptation to low gravity environments, are needed.

The hypothesis to be tested by this project is that surfaces containing copper and/or silver will inhibit biofilm formation under altered gravity regimes to a lesser extent than in 1 x g due to the fact that the interaction with the metal ions on the surface is slower because their movement around the cell is restricted to diffusion. The objective is to determine the effect and the rate, if any, of copper and/or silver surfaces on microbial growth rate, total biomass accumulation, and biofilm formation. The goal is to develop a conceptual model describing the effect of micro-gravity on biofilm formation grown on non-inhibiting surfaces as well as on metal surfaces that are potential biofilm growth inhibitors.

The approach will be to test three different microbial model systems (i.e., Escherichia coli K12, a Staphylococcus sp. isolate from the ISS, and the heavy metal resistant strain Cupriavidus metallidurans CH34) for biofilm formation on various copper- and silver-surfaces, as well as inert surfaces as controls. These surfaces differ in their antimicrobial activity based on chemical composition and/or geometric nanostructures. These surfaces will be tested for biofilm formation rates under different spaceflight relevant gravitational regimes (e.g., Moon 0.16 x g, Mars 0.38 x g, µg ISS and 1 x g control). Microbial growth will occur under optimal biofilm-inducing conditions conducted in the KUBIK incubator inside the European Drawer Rack under defined gravitational influences. Biofilm/metal surface samples and controls will be subjected to an intense analysis program, including various microbiological, genetic, molecular biological, chemical, material-science, and structural investigations. The data generated will be of immense importance for understanding the influence of µg and the ISS environment on biofilm formation as well as for the evaluation and production of improved antimicrobial additives, coating, components, surfaces and textiles for short- and long-term utilization for present and future astronaut-/robotic-associated activities in space exploration.

Research Impact/Earth Benefits:

Task Progress & Bibliography Information FY2018 
Task Progress: New project for FY2018.

Bibliography: Description: (Last Updated: 02/22/2023) 

Show Cumulative Bibliography
 
 None in FY 2018