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Feb. 2016 | Volume 4, Issue 1

STEC CAP News CONTROLLING SHIGA TOXIN-PRODUCING E. coli TO IMPROVE BEEF SAFETY

Director’s Update It is with a heavy heart that I begin this letter in remembrance of Ms. Ruth Ulmer, our STEC CAP Business Associate at the University of NebraskaLincoln, who died on November 28, 2015. Ruth was with us only eight months, but in that short span she played a significant role in our project and became a friend to Jill, myself and others here at Nebraska. She will be dearly missed. Ruth’s position has been filled by a new hire, Ms. Cara Bertlemann. Please find Cara and her contact information in the “New Faces” article in this issue of the newsletter. Welcome, Cara! In my last Director’s Update one year ago, as we entered Year 4, I made an appeal to “put your efforts into overdrive” and to prioritize, complete projects, and publish manuscripts. Now, one year later, as we enter February of Year 5, I am pleased to report that we have been successful in increasing our numbers of refereed journal publications, and I congratulate all those who have played a role in getting those done. Since the last newsletter in September, we’ve published eight refereed journal articles, have six others accepted/ in press, and several others currently under review. Although the productivity has increased, I hope we remain on an

upward slope, and continue to stress the importance of getting projects completed, papers published, and “points on the board” reported through the Piestar system (https://steccap.piestar.com). When our outside evaluator, the folks at the Office of Educational Innovation and Evaluation (OEIE) come calling on you for input (i.e., reporting of accomplishments, completion of surveys), I beg you to respond as we cannot document progress without your input.

Rod Moxley

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ear STEC CAP Nation,

Student Perspective

I would like to take a moment to congratulate those students who have graduated and gone on to start their careers, and those students who recently won awards for their STEC CAP research. My student, Zachary Stromberg, graduated with a Ph.D. in Integrative Biomedical Sciences in December after publishing several of his STEC CAP-funded research papers and will begin next week in a post-doctoral fellowship at Iowa State University doing research on… Escherichia coli, of course! We wish Zach

...continued page 5 STEC CAP Team 126D VBS Lincoln, NE 68905 PHONE: 402-472-8564 FAX: 402-472-9690

stecbeefsafety.org

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ost employees in Coles Hall can easily identify our corner of the building by the hearty laugh of our lab manager, Neil Wallace. I’d affectionately liken it to a land-mine: explosive in nature and easily triggered. Neil is rarely without a smile or a funny anecdote, and one member of our now tight-knit group that I was first introduced to as a freshly minted graduate student back in 2012. Prior to that, I was working as a fellow lab manager but in a row crops pathology lab. The experience peaked an interest in food safety and my decision to pursue graduate research in the field is what led to a chance meeting with Dr. T.G. Nagaraja in spring of 2012. Only minutes into my first sit-down with T.G., I realized that I was in the right place and in the right hands. I left Coles Hall very interested in the USDA grant objectives T.G. had described. Later that night I may have even tried (unsuccessfully) to google what a “STECK” was. Thankfully for me, T.G. took a chance on an amateur plant pathologist and later that fall I was a bona fide STEC researcher. Like all new graduate students, I was a novice immersed in a sea of experts, with the unspoken task of

...continued page 3 Inside this issue Director’s Update ...........................................1 Student Perspective........................................1 Souderton create campaign.........................2 Detection of Lipids..........................................4 STEC Conference coming ..............................6 Summer IAFP....................................................7


STEC CAP News Souderton Area High School creates food safety campaign

USDA, local businesses mentor Souderton students, create campaign

Creating an advertising campaign for the USDA, students in Souderton Area High School’s Design, Marketing, and Communications (DMC) Club from Souderton, Pennsylvania are working with mentors from local businesses and the USDA to promote food safety

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aising awareness about the potential dangers of Shiga toxin-producing Escherichia-coli (STEC) in beef and giving students a mentorship opportunity, representatives from the U.S. Department of Agriculture National Institute of Food and Agriculture (USDA NIFA) funded STEC Coordinated Agriculture Project (STEC CAP) are asking the DMC Club to create a food safety advertising campaign. According to Assistant Principal Kyle Longacre, the DMC Club formed last year and worked with the Keystone Opportunity Center. This year, the students will be creating a campaign highlighting STEC’s “potential for human health issues,” said Longacre. The process officially began in January during an informational meeting where the DMC Club met with USDA representatives and local business mentors. USDA microbiologist John Luchansky presented information about STEC for students to use in their campaigns. Students also used the meeting as an opportunity to ask questions and brainstorm ideas. Luchansky approached Longacre with this idea after learning of Souderton’s career pathway program. According to Longacre, Luchansky’s kids went through Souderton and that is how he heard about the high school’s mentorship program. Luchansky’s three daughters, Sarah, Samantha, and Stephanie, graduated in 2009, 2012, and 2014, respectively, while son Stephen is currently a sophomore at the high school. The pathways mentorship ties in with the STEC CAP grant, which also offers student internships and externships.

According to stecbeefsafety.org, the goals of the USDAsponsored STEC CAP are to help reduce health risks in beef from STEC while offering students real world experiences. Students will be creating an advertising campaign to help spread this message. USDA representatives are looking forward to working on this project with students. Luchansky is looking forward to seeing the students get “excited about food safety.”

STEC CAP News “The mentors will guide them along, and [the students will] pitch their ideas to the USDA,” said Olenick. The mentors are enthusiastic about working with students on the project. “I am excited to see what great ideas the students come up with,” said One Mile Creative editorial director Virginia Woodbury. Hochstein believes that the DMC members will introduce new perspectives. “My favorite part [of being involved with the project] is that I can learn from [the students,]” said Hochstein. New agrees. “I’m excited to see [the students’] ideas,” said New. “I’m excited to see their perspective on solving [the project] from a creative

“It’s about kids finding careers that they’re interested in and following along that career pathway,” said Marjoram. “What this does is it exposes students to several different types of careers.” According to adviser Brian Ruth, there are “so many takeaways” for students, including things like portfolio materials and exposure. Mentors are hoping that this project will help show students possible careers in the marketing or food safety fields. According to Luchansky, the program is a good way to get the “next generation of food safety professionals engaged.” The project could also open up career options in advertising and communications fields.

The student’s campaign ideas have the potential to be presented at the 2016 STEC CAP Conference.

Allebach thinks this will give students the opportunity to “learn what the process” of an advertising agency is.

“I’m hoping that there’s at least one or a few ideas that students can maybe take [to the conference in] Nebraska,” said advisor Michael Olenick. “That would be fantastic.”

This can help students realize whether they like this field of work or not.

USDA representatives will teach the students about the message they want sent to the public. According to USDA food microbiologist Anna Porto-Fett, the representatives will help to educate students on the “food safety aspect” of the project. Mentors from local advertising businesses will also help the students create their campaigns. New Idea Group president Dale New said the mentors will “guide” the students and give them background on how advertising agencies work. “[The mentors] help guide [the students] along the entire marketing process,” said Allebach Communications CEO Jamie Allebach. Students will present their ideas to the mentors. “[Mentors will provide] feedback on creative ideas,” said Allebach Communications vice president of account services Christa Ward. According to advisor Stuart Marjoram, students will be able to communicate with the mentors through the whole process to get help with their ideas.

standpoint and also from a social media standpoint.” This campaign has the potential to go from concept to reality. “It’s quite possible that the STEC grant may fund a large ad campaign based off of the ideas generated by our students,” said Longacre.

“Hopefully they’ll get a taste of whether they like the kind of work [creating a campaign] is,” said New. “Hopefully we’re opening some eyes that there are some really good and exciting and fun and rewarding careers in advertising and art.” The USDA mentors are anticipating seeing the students’ final campaigns in May.

Students can also use their involvement in the DMC Club as a mentorship for the Pathways 360 program.

“I’m really, really interested, excited, and curious about the product [the students are] going to come up with,” said Luchansky.

“[The students are] working with adult mentors,” said Longacre. “It should be an excellent learning experience for them.”

Moxley agrees. “I would hope to see some very creative effective messages that would reach targeted audiences,” said Moxley.

Junior Russell Plumb joined the club this year to gain experience working on a marketing campaign.

Students are also anticipating the outcome. “[I’m looking forward to] working with the client to produce [a campaign,]” said Plumb.

“[I am looking forward to] interactions between the client and the businesses,” said Plumb. According to Porto-Fett, the students will learn “how to develop a message” and get the “importance” of the issue across to the general public. The project will also provide club members real world experience and exposure to different careers.


STEC CAP News Discriminatory Detection of Amphiphilic Lipopolysaccharides Associated with Shiga Toxin-Producing E. coli in the Beef Chain

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etection assays largely target measurement of specific nucleic acids or proteins associated with either the pathogen or the host. An important category of biomarkers, lipidated sugars, are largely ignored because of the lack of sensitive and targeted methods for their detection. An example of a lipidated biomarker associated with Shiga toxin-producing E. coli is the virulence factor and endotoxin, lipopolysaccharide (LPS). Traditionally LPS is measured by modifications of methods that were designed for proteins such as EnzymeLinked Immunosorbent Assays (ELISAs) or by measurement of its endotoxic activity. Neither approach provides targeted detection of amphiphilic LPS, or offers significant specificity or sensitivity for serotype-specific detection in complex samples with minimal sample preparation. To overcome this limitation, the biodetection component of the STEC-CAP grant incorporates the adaptation of a tailored, novel

...continued on page 6

...Student Perspective ...continued from page 1 coming up to speed. As a part of the Objective 1 team, our research was focused on “detection”. Therefore, my initial project took me off central campus and over to the Veterinary Diagnostic Lab where I first met molecular diagnostician, Dr. Jianfa Bai. Under his expert guidance, my knowledge on the design and validation of molecular-based assays quickly synthesized, 5’ to 3’ of course. As they say though, behind every good man, there’s an even better woman. As other STEC CAP projects began to brew back at our main campus laboratory, I began working with someone quite close to Jianfa—his wife, Xiaorong Shi. “Meticulous” does not even begin to describe Xiaorong’s approach to the science she conducts in her lab and my technical skills soon began to benefit from this standard of excellence. As projects were completed in the lab, the writing process began back in the office and all of that data needed analyzed! Although I once thought statistician was an antonym for microbiologist, I quickly realized the importance of a working knowledge of statistical principles. During this time, I had the pleasure of working with an extremely gifted team of epidemiologists, including Drs. David Renter, Mike Sanderson, Natalia Cernicchiaro and Elva Cha. It quickly became evident

that the work we did in the lab would mean very little without the unique ability to make sense of all that data. The more projects I worked on, the more people I worked with, each bringing their own background, skills and expertise to the table. Critical to each and every “point on the board” were my fellow graduate students—Pragathi Shridhar, Charley Cull, Allison Mckiearnan and Diana Dewsbury. Each of them have burned the midnight oil and/or have been up well before the crack of dawn for a project, not because they were told they must, but because they each have a commitment to excellence. This is one student’s perspective during their work on the STEC CAP grant, a perspective that would not exist without the collective contributions of a group. I’m an extremely proud member of that group and I hope it shows. My advice to others: lean on your group and allow them to lean on you. Then, there will be very little you can’t accomplish, in STEC research or in life.

STEC CAP News ...Directors Update ...continued from page 1 the best in his new position. Dr. T. G. Nagaraja’s Ph.D. student, Pragathi Shridhar, won a first place award the Conference of Research Workers in Animal Diseases in December for her poster on quantification of non-O157 STEC serogroups in cattle hide samples by spiral plating and multiplex quantitative PCR methods. Congratulations, Pragathi! Congratulations to Dr. John Luchansky who in October was presented the Souderton (Pennsylvania) Area High School (SAHS) Friend of Education Award. John and Dr. Anna Porto-Fett are currently pursuing two outstanding educational opportunities with administrators at SAHS. One of these projects is featured in an article in this newsletter by Ms. Angelica Savoca, Co-Editor of the student newspaper (The Arrowhead) at SAHA, and involves the development of “STEC-Beef messages” that will ultimately be disseminated by any number of vehicles (e.g., radio spots, kiosks, banners, social media, etc.) to the “masses.” The second opportunity involves an expansion of the P-360 Program at SAHS that Assistant Principal Kyle Longacre presented at our STEC CAP Annual Meeting last summer. I applaud John and Anna for these efforts and have no doubt these efforts will be well received by the USDA-NIFA, and could become prototypes for new learning experiences at the high school level. Another exciting bit of progress I wish to mention is new diagnostic test development for non-O157 STEC, done in the laboratory of Dr. Harshini Mukundan. Harshini and her Ph.D. student, Mrs. Loreen Stromberg, have worked hard to develop a waveguide-based optical sensor for non-O157 STEC. This work is also highlighted in the present issue of the newsletter and I invite you to read all about it! I thank Loreen and Harshini for all their hard efforts and excellent progress on this work. I would like to share with you the fact that our Year 5 Continuation Funding request was submitted on November 16, 2015 and the REEport was submitted on 12-10-2015. These two items listed in some detail what the STEC CAP team accomplished in Year 4 and plans to do in Year 5, and both documents were required

for continuation funding. We are now waiting to hear back from the USDA for approval, or whether any other documentation is needed. I express my heartfelt appreciation for everyone who completed their reports and supported the continuation funding request submission in other ways. I would also like to thank Ms. Kim Gieseking, Lynne Smedjr, Carrie Snyder, and Jill Hochstein for all their excellent help in gathering budget information and their assistance in the submission of the continuation funding request. As we get further into Year 5, I will inform you about my plan and process for requesting a Year 6 no-cost extension. In closing, I would like to remind you of the next STEC CAP Annual Meeting, which will be June 14-16, 2016 in Lincoln at the Embassy Suites, and ask you to put it on your calendar. As always, please feel free to contact me, Jill, or the Executive Management Team if you have any questions or concerns. Thanks again for all you do for the STEC CAP.


STEC CAP News

STEC CAP News

...Detection ...continued from page 5

Student Loreen Stromberg, University of New Mexico, who performed most the work detailed in this article.

and patented method, developed at the Los Alamos National Laboratory for amphiphilic biomarkers, membrane insertion, for the specific detection of LPS in beef lysates. The assay exploits the amphiphilic properties of LPS, allowing for its natural association with a supported lipid bilayer, followed by detection using a fluorescently labeled antibody. This approach has been optimized on a sensitive and robust waveguidebased optical biosensor platform

sample is added to the flow cell, and if LPS (or other amphiphiles) are present, it partitions with the lipid architecture on the waveguide. Subsequent addition of a fluorescently labeled reporter antibody results in specific detection. The specificity of the assay comes from the specificity of the antibody utilized. Different serotypes of STEC and other enteric bacteria produce LPS, but they all differ with respect to their O-antigen. The team, along with Dr. Rodney Moxley at UNL and researchers at the Dana Farber Institute at Harvard University, have purified LPS associated with seven serotypes of STEC, and are in the process of isolating targeted monoclonal antibodies against LPS O-antigens, to facilitate specific detection.

Membrane Insertion: A direct method for the detection of greasy, amphiphilic biomarkers such as LPS. A waveguide is functionalized with a lipid bilayer (left most). Addition of the sample results in the association of amphiphilic biomarkers such as LPS into the supported lipid bilayer architecture. Subsequent addition of a fluorescently labeled reporter antibody facilitates sensitive detection.

that was developed at the Los Alamos National Laboratory. This platform allows for detection of fluorescently labeled biomolecules within the evanescent field of single mode planar optical waveguides, thereby minimizing non-specific interactions associated with complex samples. Using this platform, and the novel membrane insertion approach, a team of researchers from the New Mexico Consortium (Harshini Mukundan, project lead), University of New Mexico (Steven Graves, Heather Mendez and Loreen Stromberg) have developed direct detection assays for amphiphilic LPS in beef lysates. This approach utilizes a lipid bilayer functionalized waveguide as a the capture strip. The

The researchers have also validated the interaction of LPS with the lipid bilayer architecture using state of the art biophysical characterization methods (Drs. Gabriel Montano and Peter Adams, Ms. Kirstie Swingle) and statistical analysis tools (Dr. Nicolas Hengartner). The end result is a repertoire of reagents (antigens, antibodies) and methods to allow for the tailored and targeted detection of amphiphilic LPS in beef. The methods can also facilitate the detection of other amphiphilic biomarkers associated with disease.

Check us out on the Web! Visit us at: www.stecbeefsafety.org Subscribe to the listserv. Send an email to: listserv@unl.edu In the message field: subscribe stecbeefsafety This project was supported by Agriculture and Food Research Initiative Grant No. 2012-68003-30155 from the USDA National Institute of Food and Agriculture, Prevention, Detection and Control of Shiga Toxin Producing Escherichia coli (STEC) from Pre-Harvest Through Consumption of Beef Products Program –A4101.

STEC CAP Newsletter February 2016  

bi-monthly newsletter

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