Progress 09/01/20 to 08/31/22
Outputs
Target Audience:Our target audience in this reporting period was peer-scientists at the University of Nebraska-Lincoln who may be interested in using the Tecniplast ISOcage system for their research. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Training activities Held hands-on training activities in our gnotobiotic facility for our graduate students to enhance their working knowledge of how to maintain gnotobiotic conditions for their experimental animals Employed two undergraduate student workers in our facility between September 2020 and May 2021 Two of our graduate students graduated with their degrees in August 2021 (Ashley Toney earned her PhD and is now a postdoctoral associate at the University of Texas Health Sciences School of Public Health in El Paso, Texas; Jing Shao earned her MS degree and is currently applying for jobs in the food industry in Shanghai, China) Two of our undergraduate student workers graduated with their BS degrees in 2021 (Duncan Paczosa is currently a graduate student at the University of Kentucky; Nicole Nordell is currently a graduate student at the University of Nebraska-Lincoln) Hired one new graduate student (David Gomes Quintero began his PhD program in August 2021) Hired and trained four undergraduate students in Fall 2021 to work in the NGMP. One of these students worked full-time during Summer 2022, one earned their BS degree in Spring 2022 (Zeyu Xing is currently a graduate student at the University of British Columbia, Vancouver, Canada) and three have returned to work with us for the 2022-23 academic year Four new undergraduate students have been hired for Fall 2022 and are currently training to work for the 2022-23 academic year Professional development Gnotobiotic technical staff and trainees participated in an educational seminar presented by Tecniplast in April 2021 to learn about proper operation of the ISOCage system Gnotobiotic technical staff and trainees participated in an in-person, hands-on educational seminar presented by Tecniplast in January 2022 to learn about proper operation of the ISOCage system An NSF EPSCoR Track 4 grant (procedure development grant) was submitted by Dr. Jeffrey Price (NGMP Associate Director) to use the ISOcage system for experiments How have the results been disseminated to communities of interest? Provided an update to Nebraska Food for Health Center (NFHC) investigators in the NFHC newsletter that the ISOcage system will be coming to our facility Introduced the ISOcage system at the NFHC annual retreat to investigators from the University of Nebraska-Lincoln, University of Nebraska-Omaha, and University of Nebraska Medical Center What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
1. Major activities completed / experiments conducted Hired Dr. Jeffrey Price as the Associate Director of the Nebraska Gnotobiotic Mouse Program (NGMP) Completed construction of our new gnotobiotic mouse facility in November 2020; moved in and became operational during December 2020 Completed setup of the NGMP as a university-approved service center Received the ISOcage P system built to our specifications and installed it in the NGMP vivarium Researched and chose appropriate sterilant for use with the ISOcage dunk tank Hosted Tecniplast technical staff for a two-day training of NGMP staff on use of both the ISOcage racks and the ISOcage Biosafety Cabinet system Successfully tested the ability of NGMP staff to maintain germ-free status in mice over 11 weeks by having two teams of two individuals perform husbandry tasks on mice twice per week. Each team performed distinct handling methods to develop an SOP for the system. Also introduced microbiota to some cages after 5 weeks to assess ability to maintain a sterile environment between treatment groups Completed SOP document and updated our IACUC protocols accordingly for use with the ISOcage system in our vivarium Successfully performed two collaborative experiments on germ-free mice on the ISOcage rack that included every-other-day experimental manipulations of mice; all mice remained germ-free during these experiments 2. Data collected Investigated the exact configurations of the ISOcage system that were best for our needs Performed additional research to assess our operational needs for the ISOcage before placing the order Assessed cultures (both aerobic and anaerobic) of feces from mice weekly during husbandry tests for each team to determine whether mice maintained germ-free status Compared two procedures for handling supplies entering the ISOcage Biosafety cabinet for the generation of our SOP Assessed fecal cultures and performed flow cytometry analysis on spleen cells from germ-free mice treated on the ISOcage system for collaborator 3. Summary statistics and discussion of results Finalized the technical specifications of the ISOcage P housing unit, decontamination trolley, and ISOcage biosafety station prior to order, including the request for a port on the side of the biosafety station to enable connections to traditional gnotobiotic isolators and allow us to safely move mice from the isolators into the ISO cages without compromising their germ-free status After receiving the ISOcage system, we found it to function as expected, with excellent control of the mouse environment and maintenance of germ-free status in mice for at least 11 weeks (an extended timeline for most experiments performed by the NGMP) and likely longer. We also developed sterilization protocols to maintain the distinct microbial status among various treatment groups. The ISOcage will greatly expand the ability of the NGMP to complete complicated experiments with multiple treatments in germ-free or conventionalized mice. Interest in the system appears high, as inquiries for experiments using the ISOcage system have been consistently fielded since it became operational in February 2022. 4. Key outcomes or other accomplishments realized Ordered our Tecniplast ISOCage P Bioexclusion System in July 2021 ISOCage arrived in December 2021 and on-site training was provided by the vendor in January 2022 The NGMP is currently included on three grant applications to the USDA or NIH that include experiments to be performed with the ISOcage system. Two collaborators have experiments either in progress or in development using the ISOcage system
Publications
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Progress 09/01/20 to 08/31/21
Outputs
Target Audience:Our target audience in this reporting period was peer-scientists at the University of Nebraska-Lincoln who may be interested in using the Tecniplast ISOCage system for their research. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Training activities Held hands-on training activities in our gnotobiotic facility for our graduate students to enhance their working knowledge of how to maintain gnotobiotic conditions for their experimental animals Employed two undergraduate student workers in our facility between September 2020 and May 2021 Two of our graduate students graduated with their degrees in August 2021 (Ashley Toney earned her PhD and is now a postdoctoral associate at the University of Texas Health Sciences School of Public Health in El Paso, Texas; Jing Shao earned her MS degree and is currently applying for jobs in the food industry in Shanghai, China) Two of our undergraduate student workers graduated with their BS degrees in 2021 (Duncan Paczosa graduated in May 2021 is currently a graduate student at the University of Kentucky; Nicole Nordell graduated in August 2021, is currently working at the Nebraska Gnotobiotic Mouse Facility as an on-call worker, and will begin graduate school at the University of Nebraska-Lincoln in January 2022) Hired one new graduate student (David Gomes Quintero began his PhD program in August 2021) Posted an advertisement for new student workers circulated it to six campus hiring boards and departmental contacts in August 2021 to hire new students for the Fall semester Professional development Gnotobiotic technical staff and trainees participated in an educational seminar presented by Tecniplast in April 2021 to learn about proper operation of the ISOCage system How have the results been disseminated to communities of interest? Provided an update to Nebraska Food for Health Center (NFHC) investigators in the NFHC newsletter that the ISOcage system will be coming to our facility What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we will: Receive and install the IsoCage system Technical staff will receive training from Tecniplast (multiday; hands-on) Technical staff will train the student users and produce SOPs and quick guides for reference Begin experiments on the ISOcage system that further the NFHC mission
Impacts
What was accomplished under these goals?
Impact Statement: Dietary modulation of the microbiota represents an exciting new strategy for disease prevention. Unfortunately, the knowledge gap for how food components interact with the microbiome to influence health and disease is significant, and very few dietary interventions are available with well-established efficacy to justify their use. The Nebraska Food for Health Center (NFHC) is working to overcome this knowledge gap by using a systematic combination of genetic and biochemical approaches in vitro to catalogue components from food crops that have microbiome-stimulating activity. While clinical evaluation of candidate food crop components remains the primary approach for establishing efficacy, pre-clinical evaluation of candidates in germ-free mice carrying human stool microbiomes provides a powerful means to focus on candidates with the highest probability of success. Such pre-clinical screening depends on specialized resources and equipment, including access to gnotobiotic mouse models and the ability to maintain strict control over the microorganisms colonizing the mice. The purchase of a Tecniplast ISOcage P System, supported by the USDA Equipment Grant Program, will significantly expand the pre-clinical capacity of the Nebraska Gnotobiotic Mouse Facility (NGMF). The ISOcage P System is a revolutionary new mouse housing system that allows studies in mice under the strict environmental conditions necessary for microbiome research, but at higher throughput with less labor than traditional gnotobiotic isolators. The ISOcage system, in combination with the extensive expertise of the NGMF staff, will accelerate NFHC's mission to identify food molecules that modulate the gut microbiome to improve health and support USDA's goal to "improve nutritional quality... with useful biologically active food compounds to enhance human health." Objective: Purchase of a Tecniplast ISOcage P System to expand the pre-clinical capacity of the Nebraska Gnotobiotic Mouse Facility. Major activities completed / experiments conducted Hired Dr. Jeffrey Price as the Associate Director of the Nebraska Gnotobiotic Mouse Program in September 2020 Completed construction of our new gnotobiotic mouse facility in November 2020; moved in and became operational during December 2020 Completed setup of the Nebraska Gnotobiotic Mouse Program as a university-approved service center in May 2020 Data collected Investigated the exact configurations of the Isocage system that were best for our needs Performed additional research to assess our operational needs for the Isocage before placing the order Summary statistics and discussion of results Finalized the technical specifications of the ISOCage P housing unit, decontamination trolley, and ISOCage biosafety station prior to order, including the request for a port on the side of the biosafety station to enable connections to traditional gnotobiotic isolators and allow us to safely move mice from the isolators into the ISO cages without compromising their germ-free status Key outcomes or other accomplishments realized Ordered our Tecniplast ISOCage P Bioexclusion System in July 2021; estimated date of arrival is mid-December 2021 with on-site training expected to be provided by the vendor in January 2022
Publications
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