The Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD) at Kansas State University was established in 2010 to help protect the nation’s agricultural and public health sectors against a high-consequence foreign animal, emerging and zoonotic disease threats. CEEZAD has four principal missions:

Development of novel, safe, efficacious, and DIVA-compatible vaccines for prevention and control of high-impact emerging and zoonotic diseases that can be manufactured in the U.S.
Development and expansion of technologies and platforms for laboratory and point-of-need pathogen detection.
Development of models to predict high-consequence disease behavior in the U.S. to aid prevention or outbreak control.
Development of education and training programs for students, veterinarians, first responders, and researchers in high-impact animal diseases and animal emergencies.

News/Events Highlights

April 28, 2023

CEEZAD paper explores stability of SARS-CoV-2

in Biological Fluids of Animals

An article co-authored by the Director of the Center of Excellence for Emerging and Zoonotic Animal Diseases (www.ceezad.org) and the Center on Emerging and Zoonotic Infectious Diseases (CEZID; https://www.k-state.edu/cezid/) reports on new findings into the stability of the SARS-CoV-2 virus in animal fluids.

The article was co-authored by Dr. Juergen A. Richt, and published in the March 16 edition of the journal, Viruses. Other co-authors were Taeyong Kwon, Natasha N Gaudreault, Konner Cool, Chester D McDowell, and Igor Morozov, all of CEEZAD and the Richt Lab.

Since its emergence in 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has continued to evolve genetically, jump species barriers, and expand its host range. There is growing evidence of interspecies transmission including infection of domestic animals and widespread circulation in wildlife.

However, knowledge of SARS-CoV-2 stability in animal biological fluids and their role in transmission is still limited as previous studies focused on human biological fluids. This study aimed to determine the SARS-CoV-2 stability in biological fluids from three animal species, cats, sheep and white-tailed deer (WTD).

Saliva, feces, 10% fecal suspensions, and urine of cats, sheep, and WTD were mixed with a known concentration of SARS-CoV-2 and incubated under indoor and three different climatic conditions. The results show that the virus was stable for up to 1 day in the saliva of cats, sheep, and WTD regardless of the environmental conditions. The virus remained infectious for up to 6 days in feces and 15 days in fecal suspension of WTD, whereas the virus was rather unstable in feces and fecal suspensions of sheep and cats.

Researchers found the longest survival of SARS-CoV-2 in the urine of cats, sheep, and WTD. They also found that side-by-side comparison with different SARS-CoV-2 strains showed that the Alpha, Delta, and Omicron variants of concern were less stable than the ancestral Wuhan-like strain in WTD fecal suspension.

The results of the study provide valuable information for assessing the potential role of various animal biological fluids in SARS-CoV-2 transmission.

To view the full article, follow this link: https://www.mdpi.com/1999-4915/15/3/761

March 27, 2023

Call for Pilot Project Applications Center on Emerging and Zoonotic Infectious Diseases (CEZID), an NIH Center of Biomedical Research Excellence at Kansas State University

Applications due: April 20, 2023

Anticipated Decision Date: April 28, 2023

Anticipated Start Date: May 1, 2023

The KSU NIH CEZID COBRE: (www.ksu.edu/cezid) will provide investigators with support for research activities, mentoring, and access to Core Lab Services. Up to two pilot projects at up to $50,000 total direct costs starting on May 1, 2023, are anticipated. Each pilot project is for one year of initial funding and is potentially renewable for a second year, pending scientific progress and available funds. The CEZID pilot grant program is intended to enable junior and senior investigators to generate preliminary data for submission of competitive grant applications, develop new technologies, and/or achieve other goals that will better position the applicant and institution to conduct biomedical research. Applications must describe a pilot research project that fits well with the scientific theme of CEZID and must incorporate substantial use of one or more associated core labs at KSU (see: www.ksu.edu/cezid). The competition is open to all full-time faculty at any State of Kansas Regents University. Tenure-track is not required. K99/R00 holders are not eligible to lead a Pilot Project. A Pilot Project recipient cannot hold any IDeA award concurrently with a CEZID Pilot Project. Applications must be prepared in general accord with the NIH PHS 398 application guidelines (see Application process). Criteria for evaluation of Pilot Project applications are as follows:

The basic criteria for NIH grant review are found at: http://grants.nih.gov/grants/peer/peer.htm.
Additional CEZID pilot grant-specific review criteria include:

Strength of the science, and the quality and clarity of its presentation
Likelihood of the project becoming competitive for independent R01 funding
Likelihood of getting a publishable result within the one-year time frame
Relevance to the CEZID theme of emerging and/or zoonotic infectious diseases
Clear, detailed plan for utilization of one or more CEZID Core Labs
Background, experience, and career status of the applicant
Track record of past research, research grant applications, and research funding

Questions about eligibility, program details, or the appropriate inclusion within the CEZID scientific theme should be directed to Juergen Richt (jricht@vet.k-state.edu). Submit applications via email to chuncov@vet.k-state.edu no later than 5pm CDT, April 20, 2023. Applications received after this time will not be opened or reviewed. Contact Christine Huncovsky (chuncov@vet.k-state.edu) if you have questions about the submission process.

CEZID Core Facilities and Director Information

Animal Model/Pathology (AMP) Core Facility

Director: Dr. Igor Morozov (imorozov@vet.k-state.edu)

Project support is provided for animal experimental designs, training and technical support, coordination of animal experiments, gross pathology and histology of tissue samples, and development of specific reagents such as hybridomas and hyperimmune sera. A laser capture microscope is also available.

Molecular and Cellular Biology (MCB) Core Facility

Director: Dr. Juergen Richt (jricht@vet.k-state.edu)

Associate Director: Dr. Waithaka Mwangi (wmwanti@vet.k-state.edu)

The goal of the MCB Core is to assist CEZID researchers in obtaining significant research results, allowing them to effectively compete for extramural funding. Cutting edge technological support is available for flow cytometry, cell sorting, microscopy, DNA sequencing, and CRISPR applications.

General Terms and Conditions of CEZID Pilot Project Awards

Projects must make significant use of at least one Core Lab. Prospective applicants should consult with the appropriate Core Lab Director(s)/Associate Director before applying. A letter of support from the Core Director/Associate Director reflecting the feasibility of the proposed project is recommended.
Funds may be used for consumable supplies, services, or small laboratory hardware, but not for equipment (i.e., items costing > $5,000). Personnel costs are allowed, with preference given to applications that name specific individuals who are eligible to work and ready to begin no later than May 1, 2023. Summer salary is limited to a maximum of one person-month. Personnel costs may not be used to support first-year graduate students. Travel costs are limited to essential research-related travel and must be pre-approved by the CEZID PI Dr. Juergen A. Richt. Tuition costs are allowable as per standard institutional policies.
Investigators who receive CEZID pilot project support are REQUIRED to participate as fully as possible in the research meetings of the CEZID Center, as well as in seminars, workshops, and other special activities organized or sponsored by the Center.
A standard NIH progress report is required from each CEZID Pilot Project Leader by February 1, 2024 for inclusion in the COBRE annual report to the NIH.
Junior faculty recipients must have a CEZID-approved senior faculty Mentor.
- While the term Junior Faculty is not specifically defined in the RFA, the intent is that Junior Faculty member would meet the NIH Early Stage Investigator (ESI) criteria. An ESI is an individual who has not served as PD/PI for a substantial NIH independent research award, such as an R01 or U01, and has completed their terminal research degree or end of post-graduate clinical training within the past 10 years. See here for the ESI definition: https://grants.nih.gov/policy/early-investigators/index.htm
The CEZID Director may make term and budget adjustments in accordance with the intent of CEZID’s Pilot grant program and NIH policies concerning scientific overlap of projects.
By accepting CEZID funds, awardees agree to comply with any and all requirements not already mentioned that may be imposed on CEZID by NIH or other institutional authorities.

Application process

Applications must be prepared in general accord with the NIH PHS 398 application guidelines.

Applications that do not comply with these review guidelines will not be reviewed.

You are encouraged to obtain assistance from the appropriate Grant Services agency at your

University (required if a KSU participant). Suggested contacts are listed below:

􀀀 KSU Office of Research and Sponsored Projects 785-532-6195 research@k-state.eduor your assigned Grant Specialist

􀀀 KU Higuchi Biosciences Center Proposal Preparation Office 785-864-4244 or 785-864-8015 hbcgrant@ku.edu

KUMC Sponsored Programs Administration 913-588-1251 spa@kumc.edu

WSU Office of Research and Technology Transfer 316-978-3285 proposals@wichita.edu

Include the following sections, in exactly this order, compiled into a single PDF:

Face page
Project Summary
Project Narrative
Detailed budget (Budget dates are May 1, 2023- April 30, 2024)
Budget justification (use continuation page)
NIH Biosketch of applicant
NIH Biosketch of mentor (if applicable)
Other support (for applicant only)
Specific Aims (1 page) – maximum of 2 aims
Research Strategy (6 pages) – include reference to rigor of prior research and rigor and reproducibility
Vertebrate Animals – include reference to sex as a biological variable
Bibliography – limited to 1 page; use complete citations with PMIDs in NIH style
Letters of support (if applicable)

Do NOT discuss a second year of funding in your application. Additional budget information will be requested at a later date if the project is considered for funding extension.

Use 11-point Arial font with one half-inch (1/2") margins on all four sides.

Figure legends in the research strategy may use 8-point Arial font.

Letters of support from Directors of Core Labs that you will use are encouraged, but not required.

Junior Investigators must include a letter of support from their Mentor.

Submit applications as a single PDF document labeled as “PI LAST NAME” and “INSTITUTION” (e.g. RUFUS.PURINAINSTITUTE.pdf).

Submit applications no later than 5 pm CDT, April 20, 2023.

If selected for funding, applicants must provide copies of all relevant compliance approvals (IACUC, IBC, IRB) prior to the release of award funds. DO NOT submit these items at this time, but DO submit them to the compliance offices ahead of time to avoid delays in funding.

Eligible CEZID Pilot Project applications will be reviewed administratively by April 21, 2023, according to the NIH criteria and the COBRE-specific criteria described above

February 24, 2023

CEEZAD researchers assess the stability of SARS-CoV-2 variants in human biological fluids

An article co-authored by the Director of the Center of Excellence for Emerging and Zoonotic Animal Diseases (www.ceezad.org) and the Center on Emerging and Zoonotic Infectious Diseases (CEZID; https://www.k-state.edu/cezid/) assesses the ability of variants of the SARS-CoV-2 virus to survive in human biological fluids and pose threats to humans.

The article was co-authored by Dr. Juergen A. Richt, Regents and University Distinguished Professor at Kansas State University and director of CEEZAD and CEZID. It was published in the February 14 edition of Microbiology Spectrum.

Other co-authors included Taeyong Kwon, Natasha Gaudreault, Konner Cool, David Meekins and Chester McDowell, all of the Department of Diagnostic Medicine and Pathology at Kansas State University and CEEZAD.

SARS-CoV-2 is a zoonotic virus first identified in 2019, and has quickly spread worldwide. The virus is primarily transmitted through respiratory droplets from infected persons; however, the virus-laden excretions can contaminate surfaces which can serve as a potential source of infection.

Since the beginning of the pandemic, SARS-CoV-2 has continued to evolve and accumulate mutations throughout its genome leading to the emergence of variants of concern (VOCs) which exhibit increased fitness, transmissibility, and/or virulence. However, the stability of SARS-CoV-2 VOCs in biological fluids has not been thoroughly investigated.

The aim of this study was to determine and compare the stability of different SARS-CoV-2 strains in human biological fluids. Researchers demonstrate that the ancestral strain of the Wuhan-like lineage A was more stable than the Alpha VOC, and the Beta VOC in human nasal mucus and sputum. In contrast, there was no difference in stability among the three strains in dried biological fluids.

The researchers also show that the Omicron VOC was less stable than the ancestral Wuhan-like strain in nasal mucus. These studies provide insight into the effect of the molecular evolution of SARS-CoV-2 on environmental virus stability, which is important information for the development of countermeasures against SARS-CoV-2.

The findings highlight the potential risk of contaminated human biological fluids in SARS-CoV-2 transmission and contribute to the development of countermeasures against SARS-CoV-2.

The full article can be viewed by pasting this link into your browser: Ancestral Lineage of SARS-CoV-2 Is More Stable in Human Biological Fluids than Alpha, Beta, and Omicron Variants of Concern - PubMed (nih.gov)

February 24, 2023

CEEZAD Director co-authors article on a new cell line derived from Culex mosquitoes

An article co-authored by the Director of the Center of Excellence for Emerging and Zoonotic Animal Diseases (www.ceezad.org) and the Center on Emerging and Zoonotic Infectious Diseases (CEZID; https://www.k-state.edu/cezid/) reports on the establishment of a new research cell line derived from Culex mosquitoes and its permissiveness to arbovirus infection.

Other co-authors included Erin Schirtzinger of the Department of Diagnostic Medicine and Pathology at Kansas State University, William Wilson and Dana Mitzel, both of the National Bio and Agro-defense Facility (NBAF) in Manhattan, KS., and Dane Jasperson, Dustin Swanson and Barbara Drolet, of the Arthropod-borne Animal Disease Research Unit, Manhattan, KS.

The article reports on establishment of a cell line from Culex tarsalis Coquillett embryonated eggs, designated as CxTr. The cell line is heterogeneous, composed predominantly of small, round cells, and spindle-shaped cells with a doubling time of approximately 52-60 h. The identity of the cell line was verified as Cx. tarsalis by sequencing of cytochrome oxidase I. The cells were found to be free of bacteria, fungi, and mycoplasma.

The permissiveness of CxTr cells to arbovirus infection was investigated with attenuated and wildtype arboviruses from four viral families: Flaviviridae (Japanese encephalitis virus), Phenuiviridae (Rift Valley fever phlebovirus), Rhabdoviridae (vesicular stomatitis virus), and Togaviridae (Mayaro virus). All viruses were able to infect and replicate within the newly established CxTr cells.

The full article can be viewed by pasting this link in your browser: Establishment of a Culex tarsalis (Diptera: Culicidae) Cell Line and its Permissiveness to Arbovirus Infection - PubMed (nih.gov)

January 4, 2023

CEEZAD researchers advance understanding of spread of African Swine Fever Virus in Mongolia

An article co-authored by the Director of the Center of Excellence for Emerging and Zoonotic Animal Diseases (www.ceezad.org) and the Center on Emerging and Zoonotic Infectious Diseases (CEZID; https://www.k-state.edu/cezid/) sheds new light on the African Swine Fever virus which spread in domestic pigs in Mongolia.

Other co-authors included Natasha Gaudreault, Konner Cool, Jessie Trujillo, Igor Morozov, David Meekins, Chester McDowell, Dashzeveg Bold, Velmurugan Balaraman, Taeyong Kwon, Daniel Madden, Bianca L. Artiaga, Cassidy Keating, Jamie Retallick and Jayme A. Souza-Neto, all of the Department of Diagnostic Medicine and Pathology at Kansas State University and CEEZAD.

African swine fever (ASF) is an infectious viral disease caused by African swine fever virus (ASFV), that causes high mortality in domestic swine and wild boar (Sus scrofa). Currently, outbreaks are mitigated through strict quarantine measures and the culling of affected herds, resulting in massive economic losses to the global pork industry.

In 2019, an ASFV outbreak was reported in Mongolia, describing a rapidly progressing clinical disease and gross lesions consistent with the acute form of ASF.

Due to the limited information on clinical disease and viral dynamics in swine available from field observations of the Mongolian isolates, CEEZAD researchers conducted the present study to further evaluate the progression of clinical disease, virulence, and pathology of an ASFV Mongolia/2019 field isolate (ASFV-MNG19), by experimental infection of domestic pigs. Intramuscular inoculation of domestic pigs with ASFV-MNG19 resulted in clinical signs and viremia at 3 days post challenge (DPC).

Clinical disease rapidly progressed, resulting in the humane euthanasia of all pigs by 7 DPC. ASFV-MNG19 infected pigs had viremic titers of 10⁸ TCID₅₀/mL by 5 DPC and shed virus in oral secretions late in disease, as determined from oropharyngeal swabs.

Whole-genome sequencing confirmed that the ASFV-MNG19 strain used in this study was a genotype II ASFV strain highly similar to other regional strains.

The results demonstrate that ASFV-MNG19 is a virulent genotype II ASFV strain that causes acute ASF in domestic swine. The full article can be viewed by following this link: https://pubmed.ncbi.nlm.nih.gov/36560702/#:~:text=Viruses,doi%3A%2010.3390/v14122698