Professor of Biology
||Associate Professor, Department of Biology & Biotechnology Certificate Program, Ball State University, Muncie, IN
||Assistant Professor, Department of Biology & Biotechnology Certificate Program, Ball State University, Muncie, IN
| 8/99 – 5/03
||Assistant Professor, Microbiology & Molecular Biology, School of Biological Sciences, Louisiana Tech University, Ruston, LA (from which 2 years towards tenure were granted at BSU)
| 6/97 – 6/99
||Postdoctoral Associate, Department of Food Science & Technology, Mississippi State University, Starkville, MS
||Ph.D. Washington State University, Microbiology
The majority of the current research in my laboratory is focused on the search for novel antibiotics from natural environments, especially dairy farm samples such as raw milk and silage. Isolation and identification of fungi and bacterial isolates from these samples will augment their screening against ESKAPE pathogens to determine which pure cultures demonstrate antimicrobial activity. These isolates will be further described and cultured to eventually purify and identify the bioactive compounds using chromatographic and instrumental methods in chemistry. These efforts are being completed through collaboration with the Small World Initiative (SWI). This group of graduate students are just beginning this exciting work (Project #4 below).
Research underway with M.S. students and undergraduate students:
Project #1 – Bacillus spp. bacteriophage from dairy environments
Rachel H. Pittsley*1, Tykon Zubkov2, David A. Sanders3, Victoria M. Blake1, Katherine G. Kornilow1, Kyla L. Adamson4, & John L. McKillip1
1Department of Biology* & 2Department of Chemistry, Ball State University, Muncie, IN; 3Markey Center for Structural Biology, Department of Biology, Purdue University, West Lafayette, IN; 4Elanco Animal Health, Clinton, IN
Bacillus spp. are within a family of bacteria known to form spores for survival in extreme environments. The spores are resistant to temperature extremes, radiation, antibiotics, and many disinfectants. When ingested by or exposed to susceptible animal hosts, the spores can germinate and create biofilms or cause infections such as endophthalmitis, bacteremia, pneumonia, or gastroenteritis. The virulence potential makes these bacteria problematic in the food industry as pathogens and spoilers. Biological control of Bacillus spp. may be possible with bacteriophage. As a protective measure, dairy farmers could utilize a known bacteriophage after pasteurization to ensure dairy product quality and safety. The goal of this research is to isolate and characterize unknown bacteriophage found in dairy milk. By determining host range, DNA sequence, and morphology, phage candidates will be identified that may have promise as agents for biological control of Bacillus spp. of concern to dairy scientists and consumers. Raw milk, silage, tank water, manure, and soil samples were processed for phage extraction and enrichment. Plaques were subjected to DNA purification for sequencing, and for negative staining and transmission electron microscopy. Results revealed that the phage isolated in this study demonstrated a broad host range across several common Bacillus spp., established that levels of CaCl2 were highly influential for phage uptake, indicated the presence of shared amplicons between the unknown phage and Bacillus phage deep blue (NC_031056) and Bacillus phage BCP8-2 (NC_027355), revealed that glycerol precipitation was more efficient for DNA sequencing, and unveiled that the isolated phage had an icosahedral head and presence of a sheath and tail fibers, measuring approximately 100nm together and sharing characteristics with the well-known T4 bacteriophage known to infect Bacillus spp. Further host range studies, genomic analyses, and PCR will reveal if this phage is novel and a possible future biological control agent, and/or whether the phage harbors antibiotic resistance genes that could have food safety and public health implications.
M.S. Student Rachel Pittsley presents her poster on Bacillus spp. phage at the American Dairy Science Association (ADSA) conference in Cincinnati, OH during summer 2019.
Muncie (IN) high school junior Asia Wyatt presents her portion of the research on lactic acid bacteria recovery with the improved MRS formulation at the Indiana State Science and Engineering Fair, spring 2019. Asia received special recognition and an award from the Society of African American Women Chemists, and is a coauthor on this manuscript, published in 2020 in the Journal of Dairy Science.
Senior biology major Nnamdi Anene works on his portion of the lactic acid bacteria project during summer 2019. Nnamdi, who is an LSAMP (Louis Stokes Alliance for Minority Participation) Scholar, is also a coauthor on the lactic acid bacteria manuscript, published in the Journal of Dairy Science.
Project #2 – Natural Level of Lactoferrin as an Antimicrobial against S. enterica and E. coli O157:H7
Authors: E.N. Biernbaum* and J.L. McKillip
Affiliation(s): 1Department of Biology, Ball State University, Muncie, IN 47303, USA
Keywords: lactoferrin, S. enterica, E. coli O157:H7
Improper storage conditions or processing of milk leads to potential spoilage and illness. Spoilage occurs due to shifts in temperature, which allow the bacteria in milk to reproduce at an increased rate. The bacteria break down the lactose in the milk, producing lactic acid and lowering the pH. However, proteins found in raw milk have antibacterial properties, but the natural levels have not yet been studied. One of these proteins, lactoferrin, is naturally found in mammalian body fluids and has antifungal, antibacterial, and antiviral properties. Lactoferrin binds to iron and reduces the amount available for bacteria to use for growth. We are currently investigating the effectiveness of the natural level of lactoferrin as an antimicrobial on two common milk pathogens, S. enterica and E. coli O157:H7. To test these objectives, we are using a broth system to determine the optimum protein concentration and then will move into a raw milk system. The growth rate of each bacterium will be measured for 18 hours using optical density and by using the standard plate count method. We plan to see reduced density and colony counts as a result of bacterial growth inhibition due to lactoferrin. If proven to be effective, this protein could extend the shelf-life of milk, which will also reduce spoilage and the risk of contracting foodborne illnesses.
Graduate student Erika Biernbaum (M.S. 2020), here shown teaching microbiology lab in 2019, now works at Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, IA, USA
Project #3 – Fine Focus
The most innovative teaching platform I have developed is implementation of immersive learning in several courses since my last promotion. First, in 2008, with the BSU Virginia B. Ball Center for Creative Inquiry seminar entitled State of Assault, a student-produced documentary on sexual assault that won an Emmy Award in 2009, and more recently Fine Focus. Since 2013, Fine Focus has been implemented as a product-based course at Ball State University, comprised of 12-24 undergraduates each term from a variety of disciplines. Fine Focus students have developed and manage the first (and only) international digital and print journal for undergraduate microbiology research, with the American Society for Microbiology (ASM) as a community partner. The student review team in this course is responsible for making important decisions that direct the future of the journal, including learning the process of manuscript management through our double-blind peer review system using experts from our external Editorial Board. The marketing/design team is responsible for determining how to best target Fine Focus to our global audience, soliciting manuscripts, promoting Fine Focus through our website and social media, and creating an Executive Committee. This faculty-led, student-driven “immersive learning” model allows the students to be the authors of, and authorities on, their own education.
Over the last few years, Fine Focus has engaged undergraduate students in multiple STEM outreach activities, all of which offer opportunities for undergraduates to learn about and become involved in publishable undergraduate research. One of these higher profile activities is the creation of a short film entitled Faces of Science.
Faces of Science is a STEM outreach collaboration between Ball State University faculty member Dr. John McKillip (Department of Biology), his Fall 2017 Virginia B. Ball Center for Creative Inquiry class, and the University of Detroit Mercy (UDM) undergraduates who are part of an NSF-funded program called ReBUILDetroit. These students at UDM are all actively engaged in various STEM undergraduate research projects, and represent academically underrepresented student groups. This film created by Ball State University students, entitled Faces of Science, features several ReBUILDetroit undergraduates telling their individual stories about how they perceive opportunities in STEM currently, and offer reflections on their role models, advice to younger students on science, society, and the individual. Early portions of this seven-minute short film were done principally by the nine BSU biology students with post-production completion by Blake Conner (BSU Digital Corps). A link to view this inspiring film will be posted in the very near future.
Fine Focus student editors Victoria Blake and Gabriella DeValeria represent Ball State University in San Francisco by promoting and marketing Fine Focus at the ASM Microbe General meeting in June 2019.
Fine Focus implemented a local STEM outreach workshop to the Muncie (IN) Unity Center, with assistance from AmeriCorps staff of the local Second Harvest Food Bank in summer 2019. Students learned basics of microbiology theory and techniques, along with professional development opportunities.
Project #4 - Development of a multi-course CURE to transform an undergraduate microbiology curriculum and train biology majors in the process of doing science.
Doctoral student Gabriella Fluhler is beginning a dissertation research project that combines a modified CURE pedagogical model in microbiology with original research in the search for new antibiotics (Project #4).
M.S. student Madeleine Muller is underway on isolation of novel Streptomyces from dairy farm samples that may produce effective antibiotics (Project #4).
Publications (since 2015)
Church, N., & McKillip, J.L. 2021. Antibiotics and Development of Resistance. Biologia, submitted.
Brenneman, B.R., Adamson, K.A., Beer, M.R., Ho, Y., Gracias, K.S., Carstens, C.M., Biernbaum, E.N., & McKillip, J.L. 2021. Real-time screening of foods using repetitive element PCR reveals a DNA marker characteristic for enterotoxigenic Bacillus spp. Fine Focus, submitted.
Biernbaum, E.N., Gnedza, A., Akbar, S., Franklin, R., Venturelli, P. & McKillip, J.L. 2021. Lactoferrin as an antimicrobial against Salmonella enterica and Escherichia coli O157:H7 in raw milk. J. Dairy Sci. Commun., submitted
Yujun Jiang, Sihan Chen, Yueming Zhao, Xinyan Yang, Shiqian Fu, John L. McKillip, Edward M. Fox, Chaoxin Man. 2020. Multiplex Loop-Mediated Isothermal Amplification-Based Lateral Flow Dipstick for Simultaneous Detection of Three Foodborne Pathogens in Powdered Infant Formula. J. Dairy Sci.103(5):4002-4012.
Renschler, M.A., Wyatt, A., Anene, N., Robinson-Hill, R., Pickerill, E.S., Fox, N.E., Griffith, J.A., & McKillip, J.L. 2020. Using Nitrous Acid Modified MRS Medium to Selectively Isolate and Culture Lactic Acid Bacteria from Dairy Foods. J. Dairy Sci.103(2):1215-1222.
Rajabli, N., Williamson, L., Nimmer, P.S., Kelly-Worden, M., Bange, J.S., Ho, Y., & McKillip, J.L. 2019. The dangers of sublethal carvacrol exposure: increases in virulence of Bacillus cereus during endophthalmitis. International Journal of Biochemistry & Molecular Biology 9(2): 11–21.
Adamson, K.L. & McKillip, J.L. 2019. Bacillus spp. bacteriophage and the dairy environment. J. Liberal Arts & Sciences, in press.
Caffo, L., Sneed, B.L., Burcham, C., Reed, K., Hahn, N.C., Bell, S., Downham, O., Evans, M.D., Fullenkamp, C.R., Drinnon, T.K., Bishop. D., Bruns, H.A., McKillip, J.L., Sammelson, R.E., & McDowell, S.A. 2019. Simvastatin and ML141 decrease intracellular Streptococcus pyogenes infection. Current Pharmaceutical Biotechnology 20: 733-744.
Grutsch, A., P. S. Nimmer, R. H. Pittsley, K. G. Kornilow, & J. L. McKillip. 2018. Molecular Pathogenesis of Bacillus spp., with Emphasis on the Dairy Industry. Fine Focus 4(2):203-222.
McKillip, J.L. 2017. Undergraduate Research and the Global Community - Partnering with students in Iceland and Continental Europe to develop a new undergraduate microbiology research journal. Council on Undergraduate Research Quarterly 37(3):27-30.
Lan, X., Zheng, N., Zhao, S., Li, S., Zhang, Y., McKillip, J., & Wang, J. 2017. Microbiological quality of raw cow’s milk and its association with herd management practices in Northern China. J. Dairy Sci. 100(6):4294-4299. doi: 10.3168/jds.2016-11631. Epub 2017 Apr 21
Turner, D.A., Pichtel, J., Rodenas, Y., McKillip, J.L., & Goodpaster, J.V. 2015. Microbial degradation of gasoline in soil: effect of season of sampling. Forensic Sci. Intl.251:69-76.