Progress 08/01/20 to 07/31/23
Outputs
Target Audience: Beef cattle raisers, Meat processers and packers across the U.S. states. Food safety stakeholders across U.S. Policy makers associated with policies in the meat supply chain CDC personnel Research scientists investigating SARS-CoV-2 Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Postdoctoral research associates working on this projects are trained to work in BSL 3 laboratory to work with SARS-CoV-2. Provided training for the postdoctoral research associates to develop extension materials (fact sheets, short educational videos and white papers) for this project through Agrilife extension. How have the results been disseminated to communities of interest?The results from this work have been disseminated as journal papers and as pre-prints (BioRxie) for early assess of the results. Fact sheet on the importance of sanitation rotation, biofilm harbourage has been distributed through extension. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We tested our hypothesis that biofilms can act as a reservoir in protecting, harboring, and dispersing SARS-CoV-2 throughout the beef processing facility environment. We tested this hypothesis on two virus strains, SARS-CoV-2 Delta variant and Murine Hepatitis Virus (a surrogate for SARS-CoV-2). We developed mixed-species biofilm by integrating beef processing facility drain biofilm and SARS-CoV-2 delta variant or MHV on materials found in meat processing facilities (stainless steel (SS), PVC, and ceramic tiles). After exposure to the biofilm organisms for five days post-inoculation at 7°C, we conducted plaque assays to determine whether SARS-CoV-2 Delta or MHV could remain both detectable and viable. Our data showed evidence that coronaviruses can remain viable on all the surfaces tested and are also able to integrate within an environmental biofilm. Although a portion of SARS-CoV-2 and MHV was able to remain infectious after incubation with the environmental biofilm, a large reduction in plaque numbers was identified. Interestingly, we observed a 2-fold increase in the virus-environmental biofilm biovolume when compared to biofilm without virus, indicating that the biofilm bacteria both detected and reacted to the virus. Notably, our data also indicates that the multi-species biofilms from different drains locations in beef processing had varying degrees of virus viability. Biofilm from beef processing plant A integrated and had higher detectable SARS-CoV-2 than with drain biofilms from plants B and C. The bacterial component of the microbial communities and biofilm can directly or indirectly impact the outcome of infection of mammalian viruses. The virus must be stable enough to protect the viral genome from environmental exposure during transmission but malleable enough to allow disassembly and viral genome release during cell entry. To determine why SARS-CoV-2 potentially thrives in meat packaging plants, we wanted to identify how well SARS-CoV-2 RNA could remain detectable on three different meat packaging materials (plastic wrap, meat-absorbent material, and Styrofoam). From our study, we identified that SARS-CoV-2 B.1.617.2 (Delta variant) RNA was detectable on all the materials tested (plastic wrap, meat-absorbent material, and Styrofoam) after being incubated for 120 hours post-inoculation (hpi). However, there was a reduction in copy numbers following incubation on plastic wrap and meat-absorbent material, but we observed no such reduction after exposure to Styrofoam. Our results suggest that meat packaging materials such as plastic wrap, meat-absorbent material, and Styrofoam could act as a reservoir for SARS-CoV-2 to persist and spread throughout meat packaging plants, and further in the farm-to-table chain. This study investigated the viability of SARS-CoV-2 on two types of meat, ground beef, and stew-cut beef. The findings indicated that the detection of SARS-CoV-2 Delta variant RNA decreased moderately on both types of beef samples after 120 hours post-inoculation (hpi). Furthermore, there was a significant reduction in viable viral numbers after 9 hpi, but the infectious SARS-CoV-2 Delta variant was still identified after 120 hpi. Our results indicate that water evaporates more slowly from meat than from stainless steel, suggesting that the virus can survive longer on meat. This implies that meat can serve as an aqueous reservoir for the SARS-CoV-2 Delta variant, allowing it to persist and spread within meat packaging plants. The data highlight the potential risk of SARS-CoV-2 transmission through meat products and underscores the importance of implementing effective measures to prevent the spread of the virus in meat processing facilities. The goal of Objective 4 of this grant is to develop a mathematical model to predict the potential of food contamination with SARS-CoV-2 based on experimental observations. To this end, we have completed a number of research projects as described below. Microfabricated cilia: To understand viral transport by ciliary arrays in the respiratory system, we developed an engineered system of microfabricated magnetic cilia. These carpets are capable of transporting liquids and heavy solid objects in air, using a crowd-surfing effect. This effect could also be exploited for microfluidic viscosimetry and elastometry. Interestingly, we found that the flows can reverse direction depending on the ciliary elasticity. We performed Computational Fluid Dynamics (CFD) simulations to model these flows and to predict the viral transport. Bacteria transporting viruses: Next, we studied how particles such as SARS-CoV-2 viruses can be transported over large distances by active micro-swimmers such as bacteria. We again used fluid dynamics simulations to calculate the ability of such micro-swimmers to drag particles along in their wake, a process called hydrodynamic entrainment. Importantly, we found that this entrainment volume can be twenty times larger than the bacterial size, thus significantly enhancing their effective cargo capacity. This directly influences the probability of viral transmission in food-processing facilities, especially in areas where bacteria are prevalent. Viral entrainment by motile cells: To predict the potential of food contamination with SARS-CoV-2, we are studying how SARS-CoV-2 viruses can be transported by micro-organisms. Building on our results for micro-swimmers moving along straight lines, we are now investigating micro-organisms that perform biased random walks due to phototaxis and chemotaxis. First, the characteristics of these taxes are quantified experimentally in microfluidic devices subject to varying environmental conditions such as light intensity and chemical gradients, following. Using these results, we use stochastic Langevin equations to model the chemotaxis accurately in computer simulations. Subsequently, Computational Fluid Dynamics (CFD) is coupled to these simulations to quantify the hydrodynamic entrainment due to chemotactic micro-organisms. Our initial results seem to suggest that the hydrodynamic entrainment is reduced compared to linear micro-swimmers, thus suppressing the viral dispersion. Enhanced viral diffusion: Besides directed transport, SARS-CoV-2 viruses can be dispersed by micro-swimmers in a process called enhanced non-equilibrium diffusion. We developed a comprehensive model for this dispersal by generalizing Fick's laws for diffusion. Remarkably, we discovered a self-cleaning effect where surface-driven fluctuations can repel particles. Therefore, these results increase our understanding of viral dispersal driven by micro-organisms, but they could also suggest interesting strategies for making self-cleaning coating materials. Preventing viral dispersal: To prevent SARS-CoV-2 dispersal in liquid or air flows, we again used Computational Fluid Dynamics (CFD) simulations to analyze how we can drive reconfigurable flow patterns. At the macroscopic scale of cattle feedlots or rooms in food processing facilities, controlled air flows could be driven by ventilators and HVAC systems. At the microscale, these flows could be driven, for example, using the magnetic artificial cilia described above. Switching the flow velocities dynamically, we explored the possible mode structures that can emerge in these surface-actuated systems, and we used this to optimize different functions including hydrodynamic compartmentalization and chaotic mixing. These results pave the way towards restricting particle dispersal using active flow control, with applications in preventing viral spreading in food processing facilities and cattle feedlots.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2023
Citation:
eatherstone, A., Brown, A. C., & Dass, S. C. (2023). Understanding how different surfaces and environmental biofilms found in food processing plants affect the spread of COVID-19. PLOS ONE, 18(6), e0286659. https://doi.org/10.1371/journal.pone.0286659
- Type:
Journal Articles
Status:
Accepted
Year Published:
2023
Citation:
A. J. T. M. Mathijssen, M. Lisicki, V. N. Prakash, and E. J. L. Mossige, Culinary fluid mechanics and other currents in food science, Rev Mod Phys, vol. 95, no. 2, p. accepted, 2023, doi: 10.1103/RevModPhys.95.025004
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2023
Citation:
A. B. Featherstone, A. J. T. M. Mathijssen, A. Brown, and S. Chitlapilly Dass, SARS-CoV-2 RNA can remain detectable on meat packaging materials for up to five days at refrigerated temperatures, Food Microbiology, 2023.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2023
Citation:
A. B. Featherstone, A. J. T. M. Mathijssen, A. Brown, and S. Chitlapilly Dass, SARS-CoV-2 Delta Variant is Detectable and Viable on Meat Kept at Refrigerated Temperatures, Spectrum Microbiology, 2023.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2023
Citation:
A. B. Featherstone, A. J. T. M. Mathijssen, A. Brown, and S. Chitlapilly Dass, SARS-CoV-2 Delta Variant is Detectable and Viable on Meat Kept at Refrigerated Temperatures, Specturm Microbiology 2023.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2023
Citation:
Austin, B. F., Arnold, J. T. M. M., Amanda, B. & Sapna Chitlapilly, D. SARS-CoV-2 Delta Variant Remains Viable in Environmental Biofilms found in Meat Packaging Plants. bioRxiv, 2023.2006.2015.545172, doi:10.1101/2023.06.15.545172 (2023).
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Progress 08/01/21 to 07/31/22
Outputs
Target Audience:• Meat processors and packers across the U.S. states. • Food safety stakeholders across the U.S. • Research scientists investigating SARS-CoV-2 Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Postdoctoral research associates working on this projects are trained to work in BSL 3 laboratory to work with SARS-CoV-2. Provided training for the postdoctoral research associates to develop extension materials (fact sheets, short educational videos and white papers) for this project through Agrilife extension. These materials are currently under review to be distributed to our trargeted audience How have the results been disseminated to communities of interest?The results from this work have been disseminated as journal papers and as pre-prints (BioRxie) for early assess of the results. Fact sheet on the importance of sanitation rotation, biofilm harbourage has been distributed through extension. What do you plan to do during the next reporting period to accomplish the goals?• Examine the susceptibility and transmission of SARS-CoV-2 on beef cattle • Mathematical model to predict the potential of food contamination with SARS-CoV-2
Impacts
What was accomplished under these goals?
We tested our hypothesisthat biofilms can act as a reservoir in protecting, harboring, and dispersing SARS-CoV-2 throughout the beef processing facility environment. We tested this hypothesis on two virus strains, SARS-CoV-2 Delta variant and Murine Hepatitis Virus (a surrogate for SARS-CoV-2). We developed mixed-species biofilm by integrating beef processing facility drain biofilm and SARS-CoV-2 delta variant or MHV on materials found in meat processing facilities (stainless steel (SS), PVC, and ceramic tiles). After exposure to the biofilm organisms for five days post-inoculation at 7°C we conducted plaque assays to determine whether SARS-CoV-2 Delta or MHV could remain both detectable and viable. Our data showed evidence that coronaviruses can remain viable on all the surfaces tested and are also able to integrate within an environmental biofilm. Although a portion of SARS-CoV-2 and MHV was able to remain infectious after incubation with the environmental biofilm, a large reduction in plaque numbers was identified. Interestingly, we observed a 2-fold increase in the virus-environmental biofilm biovolume when compared to biofilm without virus, indicating that the biofilm bacteria both detected and reacted to the virus. Notably, our data also indicates that the multi-species biofilms from different drains locations in beef processing had varying degrees of virus viability. Biofilm from beef processing plant A integrated and had higher detectable SARS-CoV-2 than with drain biofilms from plants B and C. The bacterial component of the microbial communities and biofilm can directly or indirectly impact the outcome of infection of mammalian viruses . The virus must be stable enough to protect the viral genome from environmental exposure during transmission but malleable enough to allow disassembly and viral genome release during cell entry. Some viruses use components of the bacterial envelope to enhance virus stability. Thus, it is reasonable to consider that the different microbial communities within the processing plant can encourage or inhibit the virus particle from being part of a sanitizer-tolerant biofilm and contributing to recurrent contamination. These results indicate a complex virus-environmental biofilm interaction. There is the potential for biofilms to protect virions from disinfecting agents, which has implications for the possibility of SARS-CoV-2 prevalence within the meat processing plant environment. Also, given the highly infectious nature of SARS-CoV-2, particularly for some of the variant strains such as omicron, having even a residual level of virus present represents a severe health hazard. The increase in biofilm biovolume in response to viruses is also a concern for food safety due to the potential of the same being seen with organisms associated with food pathogens . We studied (MHV) surrogates for SARS-CoV-2 to determine viral survival on the surface of the meat, namely, stew-cut beef and ground beef, and commonly used meat packaging materials, such as plastic wrap and meat-absorbent material . From our studies, we observed the infectivity of MHV inoculated on ground beef and stew-cut beef for 48 h and saw no significant loss in infectivity for MHV from 0 to 6 h post-inoculation (hpi) (unpaired t-test) . However, beginning at nine hpi, viral infectivity steadily decreased, resulting in a 1.12-log reduction for ground beef and a 0.46-log reduction for stew-cut beef by 48 hpi. We also observed a significant persistence of MHV on meat packaging materials , with meat-absorbent material having the highest viability (3.25 × 103 ± 9.57 × 102 PFU/mL, a 0.91-log reduction after 48 hpi), followed by plastic wrap (no detectable PFU after three hpi, a 3.12-log reduction). Despite a notable reduction in infectivity, the virus could survive and remain infectious for up to 48 h at 7°C on three of the four test surfaces. Our results provide evidence that coronaviruses, such as SARS-CoV-2, could potentially survive on meat, meat-absorbent materials.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Featherstone Austin, B., Brown Amanda, C., & Chitlapilly Dass, S. (2022). Murine Hepatitis Virus, a Biosafety Level 2 Model for SARS-CoV-2, Can Remain Viable on Meat and Meat Packaging Materials for at Least 48 Hours. Microbiology Spectrum, 10(5), e01862-01822. doi: 10.1128/spectrum.01862-22
- Type:
Journal Articles
Status:
Under Review
Year Published:
2022
Citation:
Featherstone Austin, B., Brown Amanda, C., & Chitlapilly Dass, S. (2022).Understanding the Biological Factors Behind COVID-19 Outbreaks with Food Processing Plant Environmental Biofilms. PLOS One
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Arnold J. T. M. Mathijssen, Maciej Lisicki, Vivek N. Prakash, Endre J. L. Mossige, Culinary fluid mechanics and other currents in food science, (Under review) (2022)arXiv
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