The mission of IBMF is to create, hone and validate microbe detection techniques in food, water, crop plants, insects and animals, as well as study insect vectors of plant, animal and human pathogens.
Faculty scientists and graduate research assistants work in a highly specialized microbiology Biosafety Level 2 (BSL2) laboratories in the Henry Bellman Research Center. Such laboratories are required for scientists who work with exotic or potentially high-consequence pathogens. These labs have to be highly attuned to operations with respect to:
- Biosafety – maintaining the safety of lab personnel and visitors.
- Biocontainment – ensuring that high-consequence microbes do not escape.
- Biosecurity – confirming that unauthorized people do not enter the lab.
IBMF BSL2 laboratories have established specialized procedures, equipment and training. The labs are access-controlled via monitored key cards and locked doors and sealed windows to provide biosecurity. People who enter the labs must be trained, registered and escorted at all times. Every person in the lab is equipped with personal protective equipment, or PPE, such as lab coats, disposable gloves, shoe covers and hair nets, and they are trained in the proper order for donning and doffing the equipment and washing before leaving the lab. Scientists working with high-consequence microorganisms use biosafety cabinets and have access to emergency supplies and installations in the case of an accident to ensure the biosafety of all personnel. Finally, the lab is equipped with impermeable work surfaces, autoclaves, biohazard bags and plenty of disinfectants to maintain the biocontainment of organisms handled. Nothing leaves the lab that has not been washed, autoclaved or decontaminated with disinfectant.
The BSL2 laboratories are inspected annually by the Oklahoma State University Institutional Biosafety Committee (IBC), and there are regular and surprise visits by inspectors from the Centers for Disease Control (CDC) and the Animal and Plant Health Inspection Service (APHIS) of the USDA. Auditors look for documentation that there has been comprehensive training of everyone who works in the lab and specific training of visitors. The IBC, CDC and APHIS inspectors will look for documentation of inspection, maintenance and calibration of all equipment, as well as for the written protocols that will be used by the scientists as they handle high-consequence organisms.
- Comparative Analysis of the Deltocephaline Leafhopper Transcriptomes for the Elucidation
of Vector Competence Genes
- The subfamily Deltocephalinae (Hemiptera: Cicadellidae) is the largest and more diverse subfamily of Deltocephalinae .
- Most of the leafhopper species that are vectors of economically important diseases are included within this subfamily .
- Even though insect-borne plant pathogens cause serious damage to economically important crops, the mechanisms that regulate their transmission process are not completely understood .
- It is believed that the ability to effectively acquire and transmit a pathogen may be genetically regulated in the insect vectors .
- Therefore, the comparison of the transcriptomes of vector and non-vector leafhoppers can produce new insights regarding this process.
- Determining Limit of Detection of High Throughput Sequencing Diagnostic with MiFi®
High throughput sequencing (technology can be applied to plant disease diagnostics Microbe Finder MiFi ®) is an online platform for detection of plant pathogens in HTS data, eliminating pathogen isolation, bioinformatics, amplification Diagnostic sensitivity, specificity and Limit of Detection ( are crucial metrics of any diagnostic tool We present how to calculate LOD for HTS diagnostics with a statistical inference model using pathogen specific e probe matches in metagenomic data The LOD calculates the lowest levels of the target pathogen that can be reliably detected Here we used a quadratic discriminant analysis to calculate the LOD of three citrus pathogens in metagenomic HTS data The LOD assumes that positive samples have a higher e probe ‘hit x percent identity score’ and a different Normal distribution than the negative control scores LOD, formally defined as the estimated Bayes decision boundary, is computed using the mean and variance of the positive and negative groups The LOD of citrus leprosis virus C 2 citrus tristeza virus, and citrus exocortis viroid were 4 7 4 2 and 5 1 scores/ 10000 respectively, indicating when the chance of positive is 50 50 The LOD results were consistent with the RT qPCR results, however MiFi ® was found to be more sensitive In this scenario, the model is trained on a viroid and two RNA viruses, but is assumed to be true for all taxonomic groups The development of the probability model for citrus graft transmissible bacteria and a citrus specific oomycete (Phytophthora spp) is on going.
- Electropenetrograph Comparison of Male vs. Female Planthopper Probing Activity
Female Perigrinus maidis planthoppers are larger than male planthoppers. We hypothesize that females feed more on maize than do male planthoppers. The probing behavior of male vs. female planthoppers was studied using Electropenetrography (EPG). This species produced at least eight distinct waveforms, including those that represented xylem and phloem ingestion. The male planthoppers made the same waveform patterns as the females. Analysis of total probing duration, probe number, duration of xylem ingestion and duration of phloem ingestion shows that females consume more xylem and feed longer overall than do males, supporting our hypothesis that larger female bodies are supported by more feeding.
- Microbial interactions of necrophagous flies and their impact on bacterial transmission
Flies frequently interact with humans, as well as bacteria-rich environments. The roles in which flies are involved in bacterial transmission were investigated using next generation (NGS) Illumina HiseqX 150bp paired-end sequencing and bioinformatic analysis. The microbiomes of flesh flies (Sarcophagabullata) and decomposing rat tissues were compared over periods of time following rat exposure to Sarcophagidflies. Sequencing results were analyzed for presence of targeted antibiotic-resistant bacteria from the 2017 World Health Organization (WHO) Priority List using the Kraken database. A separate database was constructed using 16s sequences from bacteria of interest. Blast+/cd-hit-2d compared this database to our 16s sequences to identify our selected bacteria of interest. Spread plate dilutions were also made to observe relationships between culturable bacteria on each of the samples. Sequence comparisons revealed variance in microbiomes between different time points, as well as between the flies and rat tissue. Eleven of the WHO prioritized antibiotic-resistant pathogens were found on each of the samples. Spread plate dilutions showed larger amounts of cultured bacterial colonies on rat tissues than fly tissues, as well as an average decrease in bacterial concentrations over time. From this study, we concluded that the microbiomes of flies are influenced by exposure to bacteria-rich food sources, and are potential reservoirs for pathogenic bacteria of scientific importance.
- The Impact of Predator Non-Consumptive Effects on Transmission of a Plant Virus
It is well documented that arthropod predators play an important role in insect pest control. Along with predation, non-consumptive effects may also alter the feeding behavior of plant virus vector species, and thus alter virus transmission efficiency. We tested this hypothesis by exposing Maize Fine Streak Virus (MFSV) inoculative leafhoppers to corn plants in the presence and absence of jumping spiders or lady beetles in mesocosm and macrocosm arenas. Mesocosm trials consisted of a single corn plant enclosed in an 8” tube, with 5 inoculative leafhoppers and a predator (single jumping spider or five lady beetles). Macrocosm arenas were 10 plants placed diagonally in 2’x2’x4’ cages, with 50 inoculative leafhoppers and a caged spider propped against each plant. In mesocosm arenas, the presence of jumping spiders in close proximity to the leafhoppers led to a decrease in transmission of MFSV. However, in larger-scale macrocosm studies, there was no change in virus transmission. The presence of lady beetles did not impact virus transmission. These results suggest that certain predators in close contact with vector species could potentially limit plant-to-plant transmission of viruses.
Ochoa Corona, F., Apparatus and method for biologic sample rapid collection and recovery device, and convenient storage. Patent No.: US 9423398 B2. 2016.
L. Ma., Cold Plasma Devices for Decontamination of Foodborne Human Pathogens”. PCT/US16/43899, OSU Disclosure 2015-17
Pai, K., C. Timmons, Ramachandran, A., T. Holbrook, L. Ma., S. Madihally, and J. Jacob. 2016. “Cold Plasma Devices for Wound Care and Decontamination”. OSU Invention Disclosure No. 2017-019.
Stakeholders and Partners
- Central Intelligence Agency
- Department of Homeland Security
- Federal Bureau of Investigation
- National Security Agency
- FBI Forensics Laboratory
- Great Plains Diagnostic Network
- Homeland Security Presidential Directive (HSPD-9)
- National Plant Diagnostic Network
- Oklahoma State University
- OSU Department of Entomology & Plant Pathology
- OSU Plant Disease & Insect Diagnostic Laboratory